The document discusses strategies and considerations for the scalable purification of plasmid DNA for use in vaccine manufacturing. It covers the key upstream steps of cell harvest, lysis, and clarification. Cell harvest is typically done using centrifugation or tangential flow filtration. Lysis is usually an alkaline lysis process that must be carefully optimized to avoid damaging the plasmid DNA. Clarification can involve various pretreatment and filtration steps to reduce impurities before downstream purification. No single platform solution currently exists and processes vary between manufacturers.
Watch the presentation of this webinar here: https://bit.ly/2SWCycq
mRNA has taken center stage. Vaccines and therapeutics based on this versatile biomolecule have the potential to transform disease prevention and treatment. This webinar will explore key considerations for efficient mRNA production, starting from facility design and raw materials selection to technologies and strategies used for manufacturing.
The success of mRNA-based COVID-19 vaccines has created a significant level of interest in this versatile biomolecule for disease prevention and treatment. While production of these vaccines took place in record time, critical decisions must be made when developing novel mRNA applications to ensure manufacturability, reproducibility, and safety. This webinar will explore foundational elements of the mRNA manufacturing workflow and strategies to design the right facilities to ensure success. Topics include collaborative approaches to ensure access to high quality raw materials, application of advanced technologies for manufacturing, options for facility design and key considerations when leveraging a contract development and manufacturing partner.
In this webinar, you will learn:
• Therapeutic potential of mRNA: COVID-19 and beyond
• How mRNA manufacturing workflows and facility design have a significant impact on reproducibility and performance
• Amptec capabilities to accelerate mRNA development and manufacturing
Membrane Chromatography Solutions for Single-Use, Intensified mAb PurificationMilliporeSigma
Participate in the interactive webinar: http://bit.ly/NatrixChromMSIG
Explore our webinar library: www.emdmillipore.com/webinars
Improve productivity, flexibility, and economics of mAb purification process with intensified, single-use membrane chromatography.
Key to Successful Formulation Development for Lipid Based RNA Delivery and Va...MilliporeSigma
In this webinar, we will discuss:
• The application of RNA therapeutics and the different drug delivery routes used in the clinic.
• Design principles for developing lipids-based RNA formulations.
• Critical parameters to consider for cost effective development and consistent performance of RNA therapeutics and vaccines.
RNA therapeutics are changing the way we address diseases. Applications range from gene therapy, oncology, to vaccines for infectious diseases such as COVID-19.
The performance of RNA therapeutics critically depends on its formulation. Key decisions have to be made early on in the drug development process; choosing the appropriate drug delivery method and novel excipients. Raw material source and judicious choice of chemistry, ultimately determine the quality of novel lipid excipients which, in turn, has a big impact on the performance, reproducibility, costs, and regulatory approval timelines. This webinar will propose solutions to maximize the probability of success while formulating RNA therapeutics and vaccines.
Participate in the interactive webinar now: https://bit.ly/2xXMZlm
Explore our webinar library: www.emdmillipore.com/webinars
Single-Use Tangential Flow Filtration for Closed ProcessingMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3b7vD60
Closed processing involves use of physical barriers to separate processing fluid from the external environment. This approach reduces capital expenditures and clean room classification while accelerating time to market. This webinar will present a TFF process run in a closed mode.
Closed processing with single-use technologies is a critical enabler for efficient and robust manufacturing for novel modalities as well as continuous biomanufacturing processing. It can also reduce the dependence on classified clean rooms for traditional modalities. This approach helps to mitigate the risk of contamination by adventitious agents while enhancing operator safety.
In this presentation, we discuss the implementation of closed processing for downstream applications and present the design and performance testing of a single use manufacturing-scale tangential flow filtration system to be able to operate in both functionally and fully closed mode.
In this webinar, you will learn:
• The context of closed processing
• Differences between closed and functionally closed processing
• The drivers for adoption
• Its practical implementation to a TFF step
Unlocking the Potential of mRNA Vaccines and TherapeuticsMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3lNmkf7
The therapeutic potential of mRNA has been studied for decades and this exciting modality could potentially disrupt the biological market, in particular vaccine and novel therapies. This webinar will highlight the potential of mRNA therapies and focus on the manufacturing process's associated challenges, solutions and perspectives from synthesis to delivery.
mRNA has emerged as a promising modality for a wide range of therapeutics and vaccines and could become the break-through technology of this century. mRNA-based platform technologies could enable a more rapid response to infectious diseases, outbreaks or pandemics and allow efficient gene replacements or cancer treatments. mRNA represents a safer alternative to DNA-based therapies and the technology has recently advanced to overcome stability and efficacy challenges. Because of that, the industrialization of this technology is just in its infancy stages and bottlenecks exist around scalability, purity, and delivery which are key to establish and deliver the promise of such platform. This webinar will shed light on the potential of mRNA therapies and focus on the manufacturing process's associated challenges, solutions and perspectives from synthesis to delivery.
In this webinar, you will learn:
• The potential behind using mRNA as a therapeutic and vaccine
• The mRNA production process
• The challenges around mRNA production
• The solutions and perspectives for a robust manufacturing process
• mRNA delivery systems and their manufacturing
CAR-T Manufacturing Innovations that Work - Automating Low Volume Processes a...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3NDNIKe
Automated, fit-for-purpose tools are essential in CAR-T processing to support sustainable manufacturing of clinical and market-approved cell therapy products. This webinar will discuss how the ekko™ Acoustic Cell Processing System uses acoustic technology as a touchless approach to manipulate cells, enabling a modular tool across the CAR-T manufacturing workflow. Typical performance of templated ekko™ System processes for DMSO washout of leukapheresis material, low volume and high cell concentrate for electroporation preparation, and harvest of expanded T cells will be reviewed.
This webinar will also give an early glimpse at the ekko™ Select System for unmatched T cell selection.
In this webinar, you will:
• Uncover how the ekko™ System supports the broad industrialization of cell therapy, with particular focus on how to achieve low volume, high concentrate cell product for critical transduction and transfection steps
• Discover how ekko™ System for wash and concentrate processes throughout the cell therapy workflow achieve high cell recovery, viability, and effective residual removal
• Preview to ekko™ Select, our cell therapy selection platform, to achieve unmatched ease-of-use with direct processing from leukopaks reducing the need for preparation steps
Presented by:
Benjamin Ross-Johnsrud, Acoustic Technology Expert
Robert Scott, Mechanical Engineer III
Keeping the (Adventitious) Virus Out of the (Adeno-Associated) VirusMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/2VRylbi
How can you keep an adventitious virus from contaminating your gene therapy that is delivered by an adeno virus vector? As viral vector bioprocessing advances, regulatory requirements for viral safety will as well. Learn how to define your viral clearance strategy for AAV delivered gene therapies.
How do you define a strategy for viral clearance for a process that inherently aims at purifying a virus?
Gene delivery using AAV has received a boost from two major approvals and the nearly 300 programs in the clinic. Novel gene therapies using viral vectors enable companies to transform the lives of people living with certain rare and ultra-rare diseases where treatments are often not available currently. Amongst a multitude of challenges in viral vector bioprocessing, uncertainty in regulatory expectations is a major challenge to gene therapy developers. Regulatory requirements are evolving as the science and manufacturing matures with more stringent measures for viral safety assurance expected for future approvals.
Learn how to implement techniques for adventitious virus removal in your viral vector process; we will focus on strategies for viral clearance along your journey towards commercial readiness of AAV-based processes.
In this webinar, you will learn:
• AAV process flows and focus areas for viral safety
• Strategies for implementing viral clearance measures in bioprocessing
• Case studies and data driven approaches on log reduction values (LRV) in a viral vector process
• Best practices and evaluation roadmaps on conducting viral clearance studies
Presented by: Ratish Krishnan, Senior Strategy Consultant, Novel Modalities Bioprocessing
This presentation provides an introduction to the M Lab™ Collaboration Centers, an overview of chromatography theory, and highlights the benefits of next-generation chromatography.
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/mlab
Watch the presentation of this webinar here: https://bit.ly/2SWCycq
mRNA has taken center stage. Vaccines and therapeutics based on this versatile biomolecule have the potential to transform disease prevention and treatment. This webinar will explore key considerations for efficient mRNA production, starting from facility design and raw materials selection to technologies and strategies used for manufacturing.
The success of mRNA-based COVID-19 vaccines has created a significant level of interest in this versatile biomolecule for disease prevention and treatment. While production of these vaccines took place in record time, critical decisions must be made when developing novel mRNA applications to ensure manufacturability, reproducibility, and safety. This webinar will explore foundational elements of the mRNA manufacturing workflow and strategies to design the right facilities to ensure success. Topics include collaborative approaches to ensure access to high quality raw materials, application of advanced technologies for manufacturing, options for facility design and key considerations when leveraging a contract development and manufacturing partner.
In this webinar, you will learn:
• Therapeutic potential of mRNA: COVID-19 and beyond
• How mRNA manufacturing workflows and facility design have a significant impact on reproducibility and performance
• Amptec capabilities to accelerate mRNA development and manufacturing
Membrane Chromatography Solutions for Single-Use, Intensified mAb PurificationMilliporeSigma
Participate in the interactive webinar: http://bit.ly/NatrixChromMSIG
Explore our webinar library: www.emdmillipore.com/webinars
Improve productivity, flexibility, and economics of mAb purification process with intensified, single-use membrane chromatography.
Key to Successful Formulation Development for Lipid Based RNA Delivery and Va...MilliporeSigma
In this webinar, we will discuss:
• The application of RNA therapeutics and the different drug delivery routes used in the clinic.
• Design principles for developing lipids-based RNA formulations.
• Critical parameters to consider for cost effective development and consistent performance of RNA therapeutics and vaccines.
RNA therapeutics are changing the way we address diseases. Applications range from gene therapy, oncology, to vaccines for infectious diseases such as COVID-19.
The performance of RNA therapeutics critically depends on its formulation. Key decisions have to be made early on in the drug development process; choosing the appropriate drug delivery method and novel excipients. Raw material source and judicious choice of chemistry, ultimately determine the quality of novel lipid excipients which, in turn, has a big impact on the performance, reproducibility, costs, and regulatory approval timelines. This webinar will propose solutions to maximize the probability of success while formulating RNA therapeutics and vaccines.
Participate in the interactive webinar now: https://bit.ly/2xXMZlm
Explore our webinar library: www.emdmillipore.com/webinars
Single-Use Tangential Flow Filtration for Closed ProcessingMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3b7vD60
Closed processing involves use of physical barriers to separate processing fluid from the external environment. This approach reduces capital expenditures and clean room classification while accelerating time to market. This webinar will present a TFF process run in a closed mode.
Closed processing with single-use technologies is a critical enabler for efficient and robust manufacturing for novel modalities as well as continuous biomanufacturing processing. It can also reduce the dependence on classified clean rooms for traditional modalities. This approach helps to mitigate the risk of contamination by adventitious agents while enhancing operator safety.
In this presentation, we discuss the implementation of closed processing for downstream applications and present the design and performance testing of a single use manufacturing-scale tangential flow filtration system to be able to operate in both functionally and fully closed mode.
In this webinar, you will learn:
• The context of closed processing
• Differences between closed and functionally closed processing
• The drivers for adoption
• Its practical implementation to a TFF step
Unlocking the Potential of mRNA Vaccines and TherapeuticsMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3lNmkf7
The therapeutic potential of mRNA has been studied for decades and this exciting modality could potentially disrupt the biological market, in particular vaccine and novel therapies. This webinar will highlight the potential of mRNA therapies and focus on the manufacturing process's associated challenges, solutions and perspectives from synthesis to delivery.
mRNA has emerged as a promising modality for a wide range of therapeutics and vaccines and could become the break-through technology of this century. mRNA-based platform technologies could enable a more rapid response to infectious diseases, outbreaks or pandemics and allow efficient gene replacements or cancer treatments. mRNA represents a safer alternative to DNA-based therapies and the technology has recently advanced to overcome stability and efficacy challenges. Because of that, the industrialization of this technology is just in its infancy stages and bottlenecks exist around scalability, purity, and delivery which are key to establish and deliver the promise of such platform. This webinar will shed light on the potential of mRNA therapies and focus on the manufacturing process's associated challenges, solutions and perspectives from synthesis to delivery.
In this webinar, you will learn:
• The potential behind using mRNA as a therapeutic and vaccine
• The mRNA production process
• The challenges around mRNA production
• The solutions and perspectives for a robust manufacturing process
• mRNA delivery systems and their manufacturing
CAR-T Manufacturing Innovations that Work - Automating Low Volume Processes a...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3NDNIKe
Automated, fit-for-purpose tools are essential in CAR-T processing to support sustainable manufacturing of clinical and market-approved cell therapy products. This webinar will discuss how the ekko™ Acoustic Cell Processing System uses acoustic technology as a touchless approach to manipulate cells, enabling a modular tool across the CAR-T manufacturing workflow. Typical performance of templated ekko™ System processes for DMSO washout of leukapheresis material, low volume and high cell concentrate for electroporation preparation, and harvest of expanded T cells will be reviewed.
This webinar will also give an early glimpse at the ekko™ Select System for unmatched T cell selection.
In this webinar, you will:
• Uncover how the ekko™ System supports the broad industrialization of cell therapy, with particular focus on how to achieve low volume, high concentrate cell product for critical transduction and transfection steps
• Discover how ekko™ System for wash and concentrate processes throughout the cell therapy workflow achieve high cell recovery, viability, and effective residual removal
• Preview to ekko™ Select, our cell therapy selection platform, to achieve unmatched ease-of-use with direct processing from leukopaks reducing the need for preparation steps
Presented by:
Benjamin Ross-Johnsrud, Acoustic Technology Expert
Robert Scott, Mechanical Engineer III
Keeping the (Adventitious) Virus Out of the (Adeno-Associated) VirusMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/2VRylbi
How can you keep an adventitious virus from contaminating your gene therapy that is delivered by an adeno virus vector? As viral vector bioprocessing advances, regulatory requirements for viral safety will as well. Learn how to define your viral clearance strategy for AAV delivered gene therapies.
How do you define a strategy for viral clearance for a process that inherently aims at purifying a virus?
Gene delivery using AAV has received a boost from two major approvals and the nearly 300 programs in the clinic. Novel gene therapies using viral vectors enable companies to transform the lives of people living with certain rare and ultra-rare diseases where treatments are often not available currently. Amongst a multitude of challenges in viral vector bioprocessing, uncertainty in regulatory expectations is a major challenge to gene therapy developers. Regulatory requirements are evolving as the science and manufacturing matures with more stringent measures for viral safety assurance expected for future approvals.
Learn how to implement techniques for adventitious virus removal in your viral vector process; we will focus on strategies for viral clearance along your journey towards commercial readiness of AAV-based processes.
In this webinar, you will learn:
• AAV process flows and focus areas for viral safety
• Strategies for implementing viral clearance measures in bioprocessing
• Case studies and data driven approaches on log reduction values (LRV) in a viral vector process
• Best practices and evaluation roadmaps on conducting viral clearance studies
Presented by: Ratish Krishnan, Senior Strategy Consultant, Novel Modalities Bioprocessing
This presentation provides an introduction to the M Lab™ Collaboration Centers, an overview of chromatography theory, and highlights the benefits of next-generation chromatography.
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/mlab
Production and purification of Viral vectors for gene and cell therapy appli...Dr. Priyabrata Pattnaik
Presentation at "2016 Osong BioExcellence - Renaissance in Immunotherapy" at South Korea, an event jointly hosted by Kbio Health and Merck on 6th October 2016.
How Molecular Structure Influences Potency of a Therapeutic BiologicMerck Life Sciences
This review will give the listener an understanding of how the molecular structure, and the different ways they can be measured, influences binding and affects potency of a therapeutic biologic.
Product characterization is key to successful biological drug development. Comprehensive characterization of new therapeutic monoclonal antibodies requires a deep understanding of their structural and functional critical quality attributes (CQAs) which may impact product potency, stability and safety. Various analytical approaches can be used to characterize the effects of changes during the process of generating a biological drug.
This webinar will review some of the approaches to N-glycan profiling of monoclonal antibodies using Mass Spectrometry (MS), including Hydrogen Deuterium Exchange (HDX-MS) analytics. Using the Humira monoclonal antibody, the effect of glycosylation on the Fc-region mediated effector function was assessed with binding and CDC and ADCC activity assays. This review will give the listener an understanding of how the molecular structure, and the different ways they can be measured, influences binding and affects potency of a therapeutic biologic.
In this webinar you will learn:
- HDX-MS - when and why to use
- Glycosylation effects assessment by activity assays
Complete single-use ADC technology from development through scale-up MilliporeSigma
This webinar will talk about the benefits of single-use technologies for the manufacturing of antibody-drug conjugates and present a successful corresponding case study.
With an expected high annual growth rate of the global Antibody-drug Conjugate (ADC) market, it is essential that CMO’s have robust manufacturing platforms to ensure successful transfer to GMP production.
Single-Use Technologies provide many advantages, including improved safety, lower costs and greater flexibility. This webinar will outline the advantages of a Single Use Platform and give a case study on how it can be used to manufacture ADC projects.
In this webinar, you will learn:
● How single-use technologies can provide benefits for ADC manufacturing
● Why a solid manufacturing platform is crucial for a successful transfer to GMP production
● How a case study demonstrates the advantages of single-use equipment in a scale up to GMP production
An Integrated Approach to Ensure Viral Vector and Gene Therapy Commercial Rea...Merck Life Sciences
Come learn more about our integrated approach to ensure viral vector and gene therapy commercial readiness. We will discuss topics relating to process development for viral vector manufacturing, biosafety testing and commercial readiness.
Significant progress has been made for the use of viral vectors for gene therapy. Promising clinical trial results as well as recent FDA approval for CAR-T cell therapy to treat certain children and young adults with B-cell lymphoblastic leukemia have signaled advancements in the field. This marks a historic action, providing opportunities for new viral vector technologies to transform medicine and the way patients are treated and even cured. The need for process development for viral vector manufacturing to improve yield to meet patient demand, biosafety testing for product characterization, potency and safety and commercial readiness to accelerate therapy to-market are critically important. Here, we emphasis an integrated approach that allows our customers solutions to ensure viral vector and gene therapy commercial readiness to meet the growing market need.
In this webinar, you will learn:
● Process development advances for production scale-up of viral vectors for gene therapy
● Methods specific for viral gene therapy product characterization, purity, potency, safety and release testing
● Commercial readiness through our US and UK Centers of Excellence for viral product manufacturing
Platform Technologies to Accelerate Novel Vaccine Development and ManufacturingMilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3jmLYHu
State-of-the-art vaccine technologies are transforming vaccine development, and solutions for fast and reliable production are needed.
The vaccine industry has undergone a revolution in technology resulting in a variety of novel therapeutic platforms that accelerate development and significantly reduce the duration for process optimization and scale-up. However, challenges in maintaining efficacy and improving process robustness remain. In this presentation, we present a comparison of these novel technologies, discuss key considerations for manufacturing and share selected case studies for platforms such as virus-like-particles, viral vectors, plasmid DNA, and mRNA platform.
In this webinar, you will learn:
• Benefits of platform technologies in vaccine development
• Key considerations when deciding between platforms
• Vaccine pipeline analysis and selected case studies
Presented by:
David Loong, Ph.D, Senior Consultant, Novel Modalities Asia Pacific, Bioprocessing Strategy
Josephine Cheng, Senior Consultant, Core Modalities Asia Pacific, Bioprocessing Strategy
This presentation from IVT Network's Method Validation Conference covers required and suggested regulations and guidances for biological process specifications. It also covers dosage form considerations and specifications for other components.
Optimization of Tangential Flow Filtration Applications and Scale Up Consider...Merck Life Sciences
This presentation provides an introduction to tangential flow filtration applications for AAV and lentivirus and will review:
• Basics of tangential flow filtration (TFF)
• TFF AAV and lentivirus process overview
• Operating parameters optimization: flux-controlled microfiltration
• Scale up considerations
To learn more about this topic or collaborate with our technical experts, schedule a remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/remotevisit
This presentation reviews current trends in bioprocessing purification and includes key considerations for continuous processing and connected polishing for monoclonal antibodies. Topics include:
• Market trends and the evolution of next-generation processes
• Intensified capture processing
• Continuous virus inactivation
• Connected flow-through polishing
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/mlab
Parvovirus Filtration Best Practices - 25 Years of Hands-On ExperienceMerck Life Sciences
In this webinar, you will learn:
- how to measure filter performance and capacity,
- how to optimize filter virus removal capability,
- and avoid potential pit-falls
Detailed description:
This webinar will cover all aspects of parvovirus filtration best practices: process development/ optimization, pilot scale-up, and validation and explain the important connections between these activities. The rationale for the recommended best practices will be explained by discussing the underlying mechanisms that control filter performance.
Introduction to Tangential Flow Filtration (TFF)MilliporeSigma
This presentation provides an introduction to tangential flow filtration and reviews the following:
- TFF process basics and terminology
- TFF membrane technology
- TFF hardware, devices and systems
- Growing applications and the future
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.emdmillipore.com/mlab
Membrane Chromatography Solutions for Single-Use, Intensified mAb PurificationMerck Life Sciences
Participate in the interactive webinar: http://bit.ly/NatrixChromMRK
Improve productivity, flexibility, and economics of mAb purification process with intensified, single-use membrane chromatography.
Explore our webinar library: www.merckmillipore.com/webinars
Technology Trends in Bioprocessing PurificationMilliporeSigma
This presentation reviews current trends in bioprocessing purification and includes key considerations for continuous processing and connected polishing for monoclonal antibodies. Topics include:
• Market trends and the evolution of next-generation processes
• Intensified capture processing
• Continuous virus inactivation
• Connected flow-through polishing
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.emdmillipore.com/mlab
This presentation provides an introduction to tangential flow filtration and reviews the following:
- TFF process basics and terminology
- TFF membrane technology
- TFF hardware, devices and systems
- Growing applications and the future
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/mlab
How does the ICH Q5A revision impact viral safety strategies for biologics?MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3t7X9tg
How does the ICH Q5A revision impact viral safety strategies for biologics?
Biologics continue to grow at a fast pace. Manufactured using cell lines of human or animal origin, these are at risk of viral contamination making safety strategies critical. A comprehensive risk mitigation strategy using multiple orthogonal measures is a regulatory expectation. ICH Q5A, the globally-harmonized guideline outlines the expectations. ICH Q5A is currently being revised to address recent scientific advancements including novel therapeutic modalities, new manufacturing paradigms, updates in viral clearance applications, and alternate detection technologies. We’ll discuss the expected changes and potential impact on viral safety strategies with case studies and examples.
In this webinar, you will learn about:
• The Importance of virus testing in biologics products
• Regulatory landscape, expectations for the Q5A revision
• What's new and changing
• Examples of alternate testing schedules, impact on viral clearance
Presented by:
Manjula Aysola, Senior Regulatory Consultant
Alison Armstrong, PhD, Sr. Director, Technical and Scientific Solutions
Selecting the right aseptic filter for your process can be complicated: today’s biomanufacturer has many filter choices each offering distinct benefits. Understanding the specific needs for individual operations, in terms of flux, capacity, bioburden reduction or sterilizing performance, gamma or thermal compatibility and single or multi-use will inform decisions that have implications for the life of the process. This webinar will provide general customer guidance and explain the benefits and disadvantages of different options to help guide customers to the most appropriate filter for their operation.
In this webinar, you will learn:
- How filter design impacts performance
- Important criteria for filter selection
- New choices and options to maximize productivity for biomanufacturers
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3icKkbZ
Ensuring the safety and quality of your AAV vector is of the utmost importance. Join this webinar for a high-level overview of the regulatory requirements for AAV testing throughout the manufacturing process, as well as a more detailed look at rcAAV and infectious titer assays.
Adeno-associated virus (AAV) vectors possess a number of advantages for use in human therapy including: high titer preparations, low immunogenicity, capacity to infect a wide range of cell types, and replication deficiency. Even with these advantages, there are biosafety concerns to consider when using AAV vectors.
This webinar will discuss key regulatory considerations across the manufacturing process, from the helper/packaging plasmids through to lot release testing. We will highlight critical assays that are required and delve into specifics on replication competent AAV testing and infectious titer determination by TCID50.
In this webinar, you will learn:
• Critical biosafety considerations for AAV vectors based on the latest regulatory guidance
• How replication competent AAV testing fits into your bulk and final release testing package
• The benefits of routine and platform assays over custom assay development
Presented by:
Steven McDade, Senior Technical Specialist, Field Technology Management
Alfonso Lavorgna, Ph.D., Operations Manager, Virology Services
Implementing a Fully Single-Use, Integrated mAb Biosimilars Purification Plat...MilliporeSigma
Access the interactive recording here: https://bit.ly/2DONZaQ
Webinar summary:
1000L-scale implementation of fully connected, disposable, advanced DSP platform for next generation mAb production.
Within the biopharmaceutical industry, there is a significant shift toward higher productivity processes resulting in improved economics without compromising robustness. Therefore, integrated continuous production technologies are of greatest interest.
Next Generation Biopharmaceutical Downstream Process is a European-funded collaborative project that aims at implementing a fully integrated manufacturing platform for biosimilar mAb based on continuous chromatography, in combination with single-use disposable technologies for all unit operations of DSP on pilot/small production scale together with incorporation of advanced analytical tools.
In this webinar, you will see:
* new DSP purification template producing > 3.3 kg of mAb in 2.5 days in less than 30m²
* proof of concept for the mAb manufacturing of tomorrow
Implementing a Fully Single-Use, Integrated mAb Biosimilars Purification Plat...Merck Life Sciences
Access the interactive recording here: https://bit.ly/2DONZaQ
Webinar summary:
1000L-scale implementation of fully connected, disposable, advanced DSP platform for next generation mAb production.
Within the biopharmaceutical industry, there is a significant shift toward higher productivity processes resulting in improved economics without compromising robustness. Therefore, integrated continuous production technologies are of greatest interest.
Next Generation Biopharmaceutical Downstream Process is a European-funded collaborative project that aims at implementing a fully integrated manufacturing platform for biosimilar mAb based on continuous chromatography, in combination with single-use disposable technologies for all unit operations of DSP on pilot/small production scale together with incorporation of advanced analytical tools.
In this webinar, you will see:
* new DSP purification template producing > 3.3 kg of mAb in 2.5 days in less than 30m²
* proof of concept for the mAb manufacturing of tomorrow
Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3pCCjPF
Ensure your Adeno-Associated Virus (AAV) is safe throughout its entire drug development journey. Learn methods that will help you speed to clinic, potentially treating diseases sooner and with greater effectiveness.
The potential of gene therapies to cure previously untreatable diseases has spurred the development of novel drugs, including those based on Adeno-Associated Virus (AAV). As with all biopharmaceuticals, it is important to identify and monitor the critical quality attributes (CQAs) of these products to ensure their safety and efficacy.
In this webinar, we will present a range of orthogonal methods to understand and define the CQAs of AAV products. These include assays for the confirmation of capsid protein identity and quantity, as well as the characterization of important product-related impurities, such as aggregates. Together these methods represent a comprehensive analytical testing package to support the characterization and lot release of AAV products.
In this webinar, you will learn:
• How to identify and monitor the critical quality attributes (CQAs) of your AAV therapy
• What assays to utilize to confirm capsid protein identity and quantity
• Why you need look to product characterization to identify and remove important product-related impurities
Process Development for Cell Therapy and Viral Gene TherapyMerck Life Sciences
Today’s viral vector manufacturing processes remain challenging. Process development is a critical enabler to bring safe, effective, sustainable products to market to address patient needs. When done properly, it can reduce the timeline of the project and the cost of producing the therapeutic product.
The webinar discusses our strategies for developing lentivirus and adeno associated virus (AAV) and the impact these early decisions can have on commercial readiness.
Watch the interactive webinar now: https://bit.ly/2VplwQq
Webinar: Effective and Efficient Design of a Downstream Purification Process ...Merck Life Sciences
Register now to participate in the interactive, on-demand webinar: https://event.on24.com/wcc/r/3640127/2D3ACB02357328FE1A6C0F00083C5C06?partnerref=SlideShare
In this webinar, you will:
- Get an overview of the pDNA market
- Receive guidance for filter selection as a replacement for centrifugation
- Learn purification strategies using AEX chromatography resins and membranes
- Understand key considerations for sterile filtration
- Learn about a complete purification process flow for pDNA
Detailed description:
Plasmid DNA (pDNA) is an important component of mRNA, vaccine, and viral vector therapies. Scaling and optimizing downstream processes during manufacturing requires an in-depth knowledge of all unit operations. This webinar presents a design for a generic manufacturing template which overcomes the challenges associated with the purification of pDNA i.e high viscosity, large molecule size, shear sensitivity, and similarities with impurities. Key considerations for purification unit operations include harvest, lysis, clarification, tangential flow filtration, chromatography to sterilizing grade filtration. The webinar presents a comprehensive case study encompassing all downstream unit operations.
Webinar: Effective and Efficient Design of a Downstream Purification Process ...MilliporeSigma
Register now to participate in the interactive, on-demand webinar: https://event.on24.com/wcc/r/3640127/2D3ACB02357328FE1A6C0F00083C5C06?partnerref=SlideShare
In this webinar, you will:
- Get an overview of the pDNA market
- Receive guidance for filter selection as a replacement for centrifugation
- Learn purification strategies using AEX chromatography resins and membranes
- Understand key considerations for sterile filtration
- Learn about a complete purification process flow for pDNA
Detailed description:
Plasmid DNA (pDNA) is an important component of mRNA, vaccine, and viral vector therapies. Scaling and optimizing downstream processes during manufacturing requires an in-depth knowledge of all unit operations. This webinar presents a design for a generic manufacturing template which overcomes the challenges associated with the purification of pDNA i.e high viscosity, large molecule size, shear sensitivity, and similarities with impurities. Key considerations for purification unit operations include harvest, lysis, clarification, tangential flow filtration, chromatography to sterilizing grade filtration. The webinar presents a comprehensive case study encompassing all downstream unit operations.
Production and purification of Viral vectors for gene and cell therapy appli...Dr. Priyabrata Pattnaik
Presentation at "2016 Osong BioExcellence - Renaissance in Immunotherapy" at South Korea, an event jointly hosted by Kbio Health and Merck on 6th October 2016.
How Molecular Structure Influences Potency of a Therapeutic BiologicMerck Life Sciences
This review will give the listener an understanding of how the molecular structure, and the different ways they can be measured, influences binding and affects potency of a therapeutic biologic.
Product characterization is key to successful biological drug development. Comprehensive characterization of new therapeutic monoclonal antibodies requires a deep understanding of their structural and functional critical quality attributes (CQAs) which may impact product potency, stability and safety. Various analytical approaches can be used to characterize the effects of changes during the process of generating a biological drug.
This webinar will review some of the approaches to N-glycan profiling of monoclonal antibodies using Mass Spectrometry (MS), including Hydrogen Deuterium Exchange (HDX-MS) analytics. Using the Humira monoclonal antibody, the effect of glycosylation on the Fc-region mediated effector function was assessed with binding and CDC and ADCC activity assays. This review will give the listener an understanding of how the molecular structure, and the different ways they can be measured, influences binding and affects potency of a therapeutic biologic.
In this webinar you will learn:
- HDX-MS - when and why to use
- Glycosylation effects assessment by activity assays
Complete single-use ADC technology from development through scale-up MilliporeSigma
This webinar will talk about the benefits of single-use technologies for the manufacturing of antibody-drug conjugates and present a successful corresponding case study.
With an expected high annual growth rate of the global Antibody-drug Conjugate (ADC) market, it is essential that CMO’s have robust manufacturing platforms to ensure successful transfer to GMP production.
Single-Use Technologies provide many advantages, including improved safety, lower costs and greater flexibility. This webinar will outline the advantages of a Single Use Platform and give a case study on how it can be used to manufacture ADC projects.
In this webinar, you will learn:
● How single-use technologies can provide benefits for ADC manufacturing
● Why a solid manufacturing platform is crucial for a successful transfer to GMP production
● How a case study demonstrates the advantages of single-use equipment in a scale up to GMP production
An Integrated Approach to Ensure Viral Vector and Gene Therapy Commercial Rea...Merck Life Sciences
Come learn more about our integrated approach to ensure viral vector and gene therapy commercial readiness. We will discuss topics relating to process development for viral vector manufacturing, biosafety testing and commercial readiness.
Significant progress has been made for the use of viral vectors for gene therapy. Promising clinical trial results as well as recent FDA approval for CAR-T cell therapy to treat certain children and young adults with B-cell lymphoblastic leukemia have signaled advancements in the field. This marks a historic action, providing opportunities for new viral vector technologies to transform medicine and the way patients are treated and even cured. The need for process development for viral vector manufacturing to improve yield to meet patient demand, biosafety testing for product characterization, potency and safety and commercial readiness to accelerate therapy to-market are critically important. Here, we emphasis an integrated approach that allows our customers solutions to ensure viral vector and gene therapy commercial readiness to meet the growing market need.
In this webinar, you will learn:
● Process development advances for production scale-up of viral vectors for gene therapy
● Methods specific for viral gene therapy product characterization, purity, potency, safety and release testing
● Commercial readiness through our US and UK Centers of Excellence for viral product manufacturing
Platform Technologies to Accelerate Novel Vaccine Development and ManufacturingMilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3jmLYHu
State-of-the-art vaccine technologies are transforming vaccine development, and solutions for fast and reliable production are needed.
The vaccine industry has undergone a revolution in technology resulting in a variety of novel therapeutic platforms that accelerate development and significantly reduce the duration for process optimization and scale-up. However, challenges in maintaining efficacy and improving process robustness remain. In this presentation, we present a comparison of these novel technologies, discuss key considerations for manufacturing and share selected case studies for platforms such as virus-like-particles, viral vectors, plasmid DNA, and mRNA platform.
In this webinar, you will learn:
• Benefits of platform technologies in vaccine development
• Key considerations when deciding between platforms
• Vaccine pipeline analysis and selected case studies
Presented by:
David Loong, Ph.D, Senior Consultant, Novel Modalities Asia Pacific, Bioprocessing Strategy
Josephine Cheng, Senior Consultant, Core Modalities Asia Pacific, Bioprocessing Strategy
This presentation from IVT Network's Method Validation Conference covers required and suggested regulations and guidances for biological process specifications. It also covers dosage form considerations and specifications for other components.
Optimization of Tangential Flow Filtration Applications and Scale Up Consider...Merck Life Sciences
This presentation provides an introduction to tangential flow filtration applications for AAV and lentivirus and will review:
• Basics of tangential flow filtration (TFF)
• TFF AAV and lentivirus process overview
• Operating parameters optimization: flux-controlled microfiltration
• Scale up considerations
To learn more about this topic or collaborate with our technical experts, schedule a remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/remotevisit
This presentation reviews current trends in bioprocessing purification and includes key considerations for continuous processing and connected polishing for monoclonal antibodies. Topics include:
• Market trends and the evolution of next-generation processes
• Intensified capture processing
• Continuous virus inactivation
• Connected flow-through polishing
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/mlab
Parvovirus Filtration Best Practices - 25 Years of Hands-On ExperienceMerck Life Sciences
In this webinar, you will learn:
- how to measure filter performance and capacity,
- how to optimize filter virus removal capability,
- and avoid potential pit-falls
Detailed description:
This webinar will cover all aspects of parvovirus filtration best practices: process development/ optimization, pilot scale-up, and validation and explain the important connections between these activities. The rationale for the recommended best practices will be explained by discussing the underlying mechanisms that control filter performance.
Introduction to Tangential Flow Filtration (TFF)MilliporeSigma
This presentation provides an introduction to tangential flow filtration and reviews the following:
- TFF process basics and terminology
- TFF membrane technology
- TFF hardware, devices and systems
- Growing applications and the future
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.emdmillipore.com/mlab
Membrane Chromatography Solutions for Single-Use, Intensified mAb PurificationMerck Life Sciences
Participate in the interactive webinar: http://bit.ly/NatrixChromMRK
Improve productivity, flexibility, and economics of mAb purification process with intensified, single-use membrane chromatography.
Explore our webinar library: www.merckmillipore.com/webinars
Technology Trends in Bioprocessing PurificationMilliporeSigma
This presentation reviews current trends in bioprocessing purification and includes key considerations for continuous processing and connected polishing for monoclonal antibodies. Topics include:
• Market trends and the evolution of next-generation processes
• Intensified capture processing
• Continuous virus inactivation
• Connected flow-through polishing
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.emdmillipore.com/mlab
This presentation provides an introduction to tangential flow filtration and reviews the following:
- TFF process basics and terminology
- TFF membrane technology
- TFF hardware, devices and systems
- Growing applications and the future
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/mlab
How does the ICH Q5A revision impact viral safety strategies for biologics?MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3t7X9tg
How does the ICH Q5A revision impact viral safety strategies for biologics?
Biologics continue to grow at a fast pace. Manufactured using cell lines of human or animal origin, these are at risk of viral contamination making safety strategies critical. A comprehensive risk mitigation strategy using multiple orthogonal measures is a regulatory expectation. ICH Q5A, the globally-harmonized guideline outlines the expectations. ICH Q5A is currently being revised to address recent scientific advancements including novel therapeutic modalities, new manufacturing paradigms, updates in viral clearance applications, and alternate detection technologies. We’ll discuss the expected changes and potential impact on viral safety strategies with case studies and examples.
In this webinar, you will learn about:
• The Importance of virus testing in biologics products
• Regulatory landscape, expectations for the Q5A revision
• What's new and changing
• Examples of alternate testing schedules, impact on viral clearance
Presented by:
Manjula Aysola, Senior Regulatory Consultant
Alison Armstrong, PhD, Sr. Director, Technical and Scientific Solutions
Selecting the right aseptic filter for your process can be complicated: today’s biomanufacturer has many filter choices each offering distinct benefits. Understanding the specific needs for individual operations, in terms of flux, capacity, bioburden reduction or sterilizing performance, gamma or thermal compatibility and single or multi-use will inform decisions that have implications for the life of the process. This webinar will provide general customer guidance and explain the benefits and disadvantages of different options to help guide customers to the most appropriate filter for their operation.
In this webinar, you will learn:
- How filter design impacts performance
- Important criteria for filter selection
- New choices and options to maximize productivity for biomanufacturers
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3icKkbZ
Ensuring the safety and quality of your AAV vector is of the utmost importance. Join this webinar for a high-level overview of the regulatory requirements for AAV testing throughout the manufacturing process, as well as a more detailed look at rcAAV and infectious titer assays.
Adeno-associated virus (AAV) vectors possess a number of advantages for use in human therapy including: high titer preparations, low immunogenicity, capacity to infect a wide range of cell types, and replication deficiency. Even with these advantages, there are biosafety concerns to consider when using AAV vectors.
This webinar will discuss key regulatory considerations across the manufacturing process, from the helper/packaging plasmids through to lot release testing. We will highlight critical assays that are required and delve into specifics on replication competent AAV testing and infectious titer determination by TCID50.
In this webinar, you will learn:
• Critical biosafety considerations for AAV vectors based on the latest regulatory guidance
• How replication competent AAV testing fits into your bulk and final release testing package
• The benefits of routine and platform assays over custom assay development
Presented by:
Steven McDade, Senior Technical Specialist, Field Technology Management
Alfonso Lavorgna, Ph.D., Operations Manager, Virology Services
Implementing a Fully Single-Use, Integrated mAb Biosimilars Purification Plat...MilliporeSigma
Access the interactive recording here: https://bit.ly/2DONZaQ
Webinar summary:
1000L-scale implementation of fully connected, disposable, advanced DSP platform for next generation mAb production.
Within the biopharmaceutical industry, there is a significant shift toward higher productivity processes resulting in improved economics without compromising robustness. Therefore, integrated continuous production technologies are of greatest interest.
Next Generation Biopharmaceutical Downstream Process is a European-funded collaborative project that aims at implementing a fully integrated manufacturing platform for biosimilar mAb based on continuous chromatography, in combination with single-use disposable technologies for all unit operations of DSP on pilot/small production scale together with incorporation of advanced analytical tools.
In this webinar, you will see:
* new DSP purification template producing > 3.3 kg of mAb in 2.5 days in less than 30m²
* proof of concept for the mAb manufacturing of tomorrow
Implementing a Fully Single-Use, Integrated mAb Biosimilars Purification Plat...Merck Life Sciences
Access the interactive recording here: https://bit.ly/2DONZaQ
Webinar summary:
1000L-scale implementation of fully connected, disposable, advanced DSP platform for next generation mAb production.
Within the biopharmaceutical industry, there is a significant shift toward higher productivity processes resulting in improved economics without compromising robustness. Therefore, integrated continuous production technologies are of greatest interest.
Next Generation Biopharmaceutical Downstream Process is a European-funded collaborative project that aims at implementing a fully integrated manufacturing platform for biosimilar mAb based on continuous chromatography, in combination with single-use disposable technologies for all unit operations of DSP on pilot/small production scale together with incorporation of advanced analytical tools.
In this webinar, you will see:
* new DSP purification template producing > 3.3 kg of mAb in 2.5 days in less than 30m²
* proof of concept for the mAb manufacturing of tomorrow
Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3pCCjPF
Ensure your Adeno-Associated Virus (AAV) is safe throughout its entire drug development journey. Learn methods that will help you speed to clinic, potentially treating diseases sooner and with greater effectiveness.
The potential of gene therapies to cure previously untreatable diseases has spurred the development of novel drugs, including those based on Adeno-Associated Virus (AAV). As with all biopharmaceuticals, it is important to identify and monitor the critical quality attributes (CQAs) of these products to ensure their safety and efficacy.
In this webinar, we will present a range of orthogonal methods to understand and define the CQAs of AAV products. These include assays for the confirmation of capsid protein identity and quantity, as well as the characterization of important product-related impurities, such as aggregates. Together these methods represent a comprehensive analytical testing package to support the characterization and lot release of AAV products.
In this webinar, you will learn:
• How to identify and monitor the critical quality attributes (CQAs) of your AAV therapy
• What assays to utilize to confirm capsid protein identity and quantity
• Why you need look to product characterization to identify and remove important product-related impurities
Process Development for Cell Therapy and Viral Gene TherapyMerck Life Sciences
Today’s viral vector manufacturing processes remain challenging. Process development is a critical enabler to bring safe, effective, sustainable products to market to address patient needs. When done properly, it can reduce the timeline of the project and the cost of producing the therapeutic product.
The webinar discusses our strategies for developing lentivirus and adeno associated virus (AAV) and the impact these early decisions can have on commercial readiness.
Watch the interactive webinar now: https://bit.ly/2VplwQq
Webinar: Effective and Efficient Design of a Downstream Purification Process ...Merck Life Sciences
Register now to participate in the interactive, on-demand webinar: https://event.on24.com/wcc/r/3640127/2D3ACB02357328FE1A6C0F00083C5C06?partnerref=SlideShare
In this webinar, you will:
- Get an overview of the pDNA market
- Receive guidance for filter selection as a replacement for centrifugation
- Learn purification strategies using AEX chromatography resins and membranes
- Understand key considerations for sterile filtration
- Learn about a complete purification process flow for pDNA
Detailed description:
Plasmid DNA (pDNA) is an important component of mRNA, vaccine, and viral vector therapies. Scaling and optimizing downstream processes during manufacturing requires an in-depth knowledge of all unit operations. This webinar presents a design for a generic manufacturing template which overcomes the challenges associated with the purification of pDNA i.e high viscosity, large molecule size, shear sensitivity, and similarities with impurities. Key considerations for purification unit operations include harvest, lysis, clarification, tangential flow filtration, chromatography to sterilizing grade filtration. The webinar presents a comprehensive case study encompassing all downstream unit operations.
Webinar: Effective and Efficient Design of a Downstream Purification Process ...MilliporeSigma
Register now to participate in the interactive, on-demand webinar: https://event.on24.com/wcc/r/3640127/2D3ACB02357328FE1A6C0F00083C5C06?partnerref=SlideShare
In this webinar, you will:
- Get an overview of the pDNA market
- Receive guidance for filter selection as a replacement for centrifugation
- Learn purification strategies using AEX chromatography resins and membranes
- Understand key considerations for sterile filtration
- Learn about a complete purification process flow for pDNA
Detailed description:
Plasmid DNA (pDNA) is an important component of mRNA, vaccine, and viral vector therapies. Scaling and optimizing downstream processes during manufacturing requires an in-depth knowledge of all unit operations. This webinar presents a design for a generic manufacturing template which overcomes the challenges associated with the purification of pDNA i.e high viscosity, large molecule size, shear sensitivity, and similarities with impurities. Key considerations for purification unit operations include harvest, lysis, clarification, tangential flow filtration, chromatography to sterilizing grade filtration. The webinar presents a comprehensive case study encompassing all downstream unit operations.
Straight to the Point: Reaching Clinical Stage Development with a CHOZN® Cell...MilliporeSigma
Participate in the interactive webinar: http://bit.ly/CHOZNWebinar
In this case study, we will present how we support our clients thanks to advantages provided by the CHOZN® Cell Line, and a specific strategy for clone selection where semi-automation and pool selection are leveraged, to get upstream right first time.
Explore our webinar library: www.emdmillipore.com/webinars
Straight to the Point: Reaching Clinical Stage Development with a CHOZN® Cell...Merck Life Sciences
Participate in the interactive webinar: http://bit.ly/CHOZNWebinar
In this case study, we will present how we support our clients thanks to advantages provided by the CHOZN® Cell Line, and a specific strategy for clone selection where semi-automation and pool selection are leveraged, to get upstream right first time.
Explore our webinar library: www.merckmillipore.com/webinars
CAR-T Manufacturing Innovations that Work - Automating Low Volume Processes a...MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3NDNIKe
Automated, fit-for-purpose tools are essential in CAR-T processing to support sustainable manufacturing of clinical and market-approved cell therapy products. This webinar will discuss how the ekko™ Acoustic Cell Processing System uses acoustic technology as a touchless approach to manipulate cells, enabling a modular tool across the CAR-T manufacturing workflow. Typical performance of templated ekko™ System processes for DMSO washout of leukapheresis material, low volume and high cell concentrate for electroporation preparation, and harvest of expanded T cells will be reviewed.
This webinar will also give an early glimpse at the ekko™ Select System for unmatched T cell selection.
In this webinar, you will:
• Uncover how the ekko™ System supports the broad industrialization of cell therapy, with particular focus on how to achieve low volume, high concentrate cell product for critical transduction and transfection steps
• Discover how ekko™ System for wash and concentrate processes throughout the cell therapy workflow achieve high cell recovery, viability, and effective residual removal
• Preview to ekko™ Select, our cell therapy selection platform, to achieve unmatched ease-of-use with direct processing from leukopaks reducing the need for preparation steps
Presented by:
Benjamin Ross-Johnsrud, Acoustic Technology Expert
Robert Scott, Mechanical Engineer III
Garvan Flow Cytometry Instrument talk. Given by Dr. Katharina Winnemöller, Research and Clinical Sales Manager (NSW, ACT) at the Garvan Instituteof Medical Research on the 15th October 2010.
AutoMacs Pro magnetic Cell Seapartion - "the faster way to ultra-pure, viable cells"
Delivering More Efficient Therapeutic Protein Expression Systems Through Cell...MilliporeSigma
Historically cell line performance has been enhanced through media, feed and process optimization, primarily through trying to meet the basic nutritional requirements of the cells so that they can sustain high growth and productivity throughout the production runs.
However, the omics (genomics, transciptomics and metabolomics) era, sequencing of the CHO genome and enhancements in genome editing technologies over the past several years have enabled scientists to take a more direct route in cell line optimization through the modification of specific genes that have direct implications on cell culture performance, protein quality attributes and upstream and downstream manufacturing processes. These targets include but are not limited to genes that may be involved in cell cycle regulation, cellular metabolism, cellular transcription and translation, the secretory pathway and protein glycosylation or other post-translational modifications.
In this webinar we will discuss specific genetic modifications that have been made to CHO cell lines and how these modifications can lead to more efficient expression systems.
Delivering More Efficient Therapeutic Protein Expression Systems Through Cell...Merck Life Sciences
Historically cell line performance has been enhanced through media, feed and process optimization, primarily through trying to meet the basic nutritional requirements of the cells so that they can sustain high growth and productivity throughout the production runs.
However, the omics (genomics, transciptomics and metabolomics) era, sequencing of the CHO genome and enhancements in genome editing technologies over the past several years have enabled scientists to take a more direct route in cell line optimization through the modification of specific genes that have direct implications on cell culture performance, protein quality attributes and upstream and downstream manufacturing processes. These targets include but are not limited to genes that may be involved in cell cycle regulation, cellular metabolism, cellular transcription and translation, the secretory pathway and protein glycosylation or other post-translational modifications.
In this webinar we will discuss specific genetic modifications that have been made to CHO cell lines and how these modifications can lead to more efficient expression systems.
Employing Innovative Platform Manufacturing and Biosafety Testing for your Ge...Merck Life Sciences
Watch the webinar here: https://event.on24.com/wcc/r/2003970/F5AFA4FE6C60AD00635D4D15BADB5D8E?partnerref=slideshare
As gene therapies and gene-modified cell therapies show increasing promise, the need for innovative and proficient viral vector manufacturing continues to grow. Concurrently, increased regulatory guidance governing the manufacturing and testing of viral vectors adds complexity and increases the timelines to successfully produce high-quality virus ready for clinical use.
This webinar will address how the implementation of both manufacturing templates and platform characterization and safety assays can increase the likelihood of success in process validation and reduce risk in the timeline to commercialization for your gene therapy product. Using adeno-associated virus (AAV) as a case study, we will demonstrate how our validated, templated process for production can reduce the need for qualification inherent in niche manufacturing workflows and anticipate forthcoming needs for process performance qualification. This webinar will also highlight benefits from a new, platform assay offering for characterization and safety testing of AAV. Because these assays are pre-qualified, they reduce the variability inherent in assay validation and subsequently the time needed to establish readiness for regulatory compliance.
While these developments increase the standardization across the manufacturing and testing workflows, they remain flexible to clients' needs and are created to be scalable and as future-proof as possible, allowing for adaptability as the regulatory landscape of gene therapies evolves.
In this webinar, you will learn:
● The unit operations in AAV manufacturing that are ideal for templating
● How the manufacturing workflow can be targeted to reduce variability in testing and improve readiness for commercial production
● How platform assays can ease the burden of assay qualification and improve overall commercialization timelines
Employing Innovative Platform Manufacturing and Biosafety Testing for your Ge...MilliporeSigma
Watch the webinar here: https://event.on24.com/wcc/r/2003970/F5AFA4FE6C60AD00635D4D15BADB5D8E?partnerref=slideshare
As gene therapies and gene-modified cell therapies show increasing promise, the need for innovative and proficient viral vector manufacturing continues to grow. Concurrently, increased regulatory guidance governing the manufacturing and testing of viral vectors adds complexity and increases the timelines to successfully produce high-quality virus ready for clinical use.
This webinar will address how the implementation of both manufacturing templates and platform characterization and safety assays can increase the likelihood of success in process validation and reduce risk in the timeline to commercialization for your gene therapy product. Using adeno-associated virus (AAV) as a case study, we will demonstrate how our validated, templated process for production can reduce the need for qualification inherent in niche manufacturing workflows and anticipate forthcoming needs for process performance qualification. This webinar will also highlight benefits from a new, platform assay offering for characterization and safety testing of AAV. Because these assays are pre-qualified, they reduce the variability inherent in assay validation and subsequently the time needed to establish readiness for regulatory compliance.
While these developments increase the standardization across the manufacturing and testing workflows, they remain flexible to clients' needs and are created to be scalable and as future-proof as possible, allowing for adaptability as the regulatory landscape of gene therapies evolves.
In this webinar, you will learn:
● The unit operations in AAV manufacturing that are ideal for templating
● How the manufacturing workflow can be targeted to reduce variability in testing and improve readiness for commercial production
● How platform assays can ease the burden of assay qualification and improve overall commercialization timelines
Investing in Process Development for Increased MSC Production in Stirred Tank...MilliporeSigma
Interested in developing a robust cell therapy manufacturing platform? In this webinar we will share information in the form of case studies that highlight strategies to optimize your cell therapy production process.
Industry trends in regenerative medicine highlight a critical need for closed cell culture systems that support scalable manufacturing of adherent cell therapies. Typical static in vitro culture methods, however, are often too cumbersome and inefficient to support commercial scale production of mesenchymal stem/stromal cells (MSCs). Single-use stirred tank bioreactor systems are a platform that can address this limitation and have been proven effective for microcarrier-based production of adherent cell therapies. Implementation of optimized process control strategies for parameters such as dissolved oxygen (DO) and agitation rate are key to making an efficient transition from planar culture to stirred tank bioreactors. Herein, a stepwise approach to process development for MSC expansion in a small-scale single-use bioreactor is presented. Case studies focus on strategies to optimize DO control and agitation rates for bone marrow derived MSCs in microcarrier culture, highlighting improvements in process efficiency. In the first case study, the impact different gassing methods have on DO control and whether hypoxic growth conditions affect MSC function are examined. The second case study demonstrates the application of Zwietering’s equation for suspension of solids to overcome scaling challenges often associated with microcarrier culture in stirred tanks. Strategies to further improve the seeding process for bioreactor culture will also be reviewed. Identifying optimal seeding and process control strategies for microcarrier-based bioreactor expansion of adherent cells is paramount for the development of robust cell therapy manufacturing platforms.
In this webinar, you will learn about:
· Process development approaches for production scale-up of mesenchymal stem cells (MSCs)
· Implementing single-use, closed systems for manufacturing cell therapies
· Case studies focusing on strategies to optimize DO control and agitation rates for microcarrier-based cultures
Investing in Process Development for Increased MSC Production in Stirred Tank...Merck Life Sciences
Interested in developing a robust cell therapy manufacturing platform? In this webinar we will share information in the form of case studies that highlight strategies to optimize your cell therapy production process.
Industry trends in regenerative medicine highlight a critical need for closed cell culture systems that support scalable manufacturing of adherent cell therapies. Typical static in vitro culture methods, however, are often too cumbersome and inefficient to support commercial scale production of mesenchymal stem/stromal cells (MSCs). Single-use stirred tank bioreactor systems are a platform that can address this limitation and have been proven effective for microcarrier-based production of adherent cell therapies. Implementation of optimized process control strategies for parameters such as dissolved oxygen (DO) and agitation rate are key to making an efficient transition from planar culture to stirred tank bioreactors. Herein, a stepwise approach to process development for MSC expansion in a small-scale single-use bioreactor is presented. Case studies focus on strategies to optimize DO control and agitation rates for bone marrow derived MSCs in microcarrier culture, highlighting improvements in process efficiency. In the first case study, the impact different gassing methods have on DO control and whether hypoxic growth conditions affect MSC function are examined. The second case study demonstrates the application of Zwietering’s equation for suspension of solids to overcome scaling challenges often associated with microcarrier culture in stirred tanks. Strategies to further improve the seeding process for bioreactor culture will also be reviewed. Identifying optimal seeding and process control strategies for microcarrier-based bioreactor expansion of adherent cells is paramount for the development of robust cell therapy manufacturing platforms.
In this webinar, you will learn about:
· Process development approaches for production scale-up of mesenchymal stem cells (MSCs)
· Implementing single-use, closed systems for manufacturing cell therapies
· Case studies focusing on strategies to optimize DO control and agitation rates for microcarrier-based cultures
Plasmid Manufacturing Service from GenScript ProBioGenScript ProBio
GenScript ProBio offers the best Plasmid Manufacturing Service and employs a GMP-compliant plasmid production process that allows customers to replicate DNA used in experiments with minimal additional effort. By employing this process, Genscript can provide plasmids produced at the highest quality standards. For more information, visit our website. https://www.genscriptprobio.com/gct-proplasmid.html
Applied StemCell Inc’s MAPK genomic DNA (gDNA) reference standards represent biologically-relevant controls that can be directly incorporated into your sample processing workflows in order to optimize your protocols, evaluate assay sensitivity and specificity, and analyze the impact of workflow changes on downstream analysis. They represent ideal materials for both assay development and routine monitoring of assay performance.
The MAPK Genomic DNA Reference Standards are extracted from ASC’s panel of isogenic MAPK mutation cell lines with 50 recurrent pathway-activating mutations in the EGFR, KRAS and BRAF genes, based on data from the Sanger Institute’s COSMIC database.
Key Features of the MAPK Series gDNA Reference Standards:
Most comprehensive MAPK mutation panel on the market
Well-characterized colorectal cancer cells lines: EGFR (RKO), KRAS (RKO), BRAF (HCT116)
Paired, isogenic wild-type cell lines to serve as an ideal control
Footprint-free, homozygous mutations
Reference cell lines are expanded from single-cells, ensuring maximum homogeneity
Available in multiple formats, including slides, scrolls, and full FFPE blocks
Why Use Reference Materials (DNA: Reference standadrds provide a consistent and reliable resource for evaluating and optimizing various stages in your sample processing workflow. Whether you’re starting from DNA extraction, assay design, or library preparation, our reference materials can help you to identify and eliminate sources of variability within your protocols.
@AppliedStemCell offers validated, cellular reference standards for direct incorporation into sample processing workflows or quality control processes.
Highlights:
Overview of molecular reference materials
Workflow and QC for ONCOREF™ cell line generation (#CRISPR)
Advantages of CRISPR-engineered molecular reference standards
Applications of reference materials in assay development
Q & A
#sangersequencing #ngs
Similar to Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing (20)
The Viscosity Reduction Platform: Viscosity-reducing excipients for improveme...Merck Life Sciences
Protein viscosity is a major challenge in preparing highly concentrated protein formulations suitable for subcutaneous injection. Recently, the Viscosity Reduction Platform (VRP) was introduced and its technical key features and benefits for formulations were discussed. However, highly viscous solutions do not only pose a challenge when administering a drug to a patient, they can also impose technical limitations in the manufacturing process.
This white paper evaluates the effect of the excipients in the Viscosity Reduction Platform on ultrafiltration processes used to produce a highly concentrated formulation of a monoclonal antibody (mAb). Two filtration methods are demonstrated in this work.
Find more information about the Viscosity Reduction Platform on our website: https://www.sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
Use of Excipients in Downstream Processing to Improve Protein PurificationMerck Life Sciences
Excipients are used to improve the stability of protein-based therapeutics by protecting the protein against a range of stress conditions such as temperature changes, pH changes, or agitation. Similar stresses are applied to proteins during downstream purification. Shifts in pH during Protein A chromatography, subsequent incubations at low pH for virus inactivation, and changes in conductivity in ion exchange chromatography can lead to aggregation, fragmentation, or other chemical modifications of the therapeutic protein. Given the potential impact on the protein’s structural integrity, there is a need for approaches to reduce the risk presented by the conditions during downstream processing. For example, integration of a solution to prevent aggregation of proteins would be a more efficient strategy than implementing steps to remove multimeric forms.
This white paper highlights the results from a recent paper by Stange et. al., in which protein stabilizing excipients such as polyols, sugars, and polyethylene glycol (PEG4000) were used as buffer system additives. Effect of the excipients on elution patterns, stabilization of the monomer antibody, host-cell protein removal, virus inactivation rates and binding capacity of cation exchange chromatography were explored.
Exploring the protein stabilizing capability of surfactants against agitation...Merck Life Sciences
Agitation of therapeutic protein solutions during manufacturing, shipping and handling is one of the major initiators for protein aggregation and particle formation during the life history of a protein drug. Adsorption of protein molecules to liquid-air interfaces leads to the formation of highly concentrated protein surface films. The rupture of these protein films due to various mechanical processes can then result in the appearance of protein aggregates and particles in the bulk solution phase.
One technique to stabilize proteins against stress induced by liquid-air interfaces is the use of non-ionic surfactants. About 91% of antibody formulations commercially available in 2021 contained a surfactant. Polysorbate 20 and 80, composed of a hydrophilic polyoxyethylene sorbitan and hydrophobic fatty acid esters, made up the largest part being employed in 87% of said formulations.
Despite their frequent use in parenteral drug products, concerns have been raised for decades about the application of polysorbates as surfactants in biopharmaceutical formulations. Autoxidation of polysorbate, caused by residual peroxides in polysorbates, can damage the proteins and can further drive the oxidative degradation of polysorbate. Chemical and enzymatic hydrolysis of polysorbate may lead to the formation of free fatty acid particles, which may become visible; and both mechanisms eventually lead to the reduction in polysorbate concentration. Therefore, the purpose of the current study was to compare various molecules for their capabilities to reduced agitation-induced protein aggregation and particle formation; and furthermore, investigate their underlying protein stabilizing mechanisms.
The Viscosity Reduction Platform: Viscosity Reducing Excipients for Protein F...Merck Life Sciences
Protein viscosity is one of the major obstacles in preparing highly concentrated protein formulations suitable for subcutaneous injection.
This whitepaper examines how combining an amino acid with a second viscosity-reducing excipient circumvents adverse effects on protein stability and improves viscosity-reducing capacity.
To find more information about the Viscosity Reduction Platform, please visit our website: https://sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
Characterization of monoclonal antibodies and Antibody drug conjugates by Sur...Merck Life Sciences
Watch the presentation of this webinar: https://bit.ly/3Pjpjvr
Highlights of this webinar:
- Surface plasmon resonance as a powerful tool for biologic characterization including mAbs and ADCs.
- SPR allows rapid binding analysis in real time without using labels for SARS-CoV-2 receptor binding domain mutations.
- Kinetic data is indicative of possible neutralizing activity allowed assessment of neutralizing ability of therapeutic monoclonal antibodies.
- The application can provide preliminarily efficacy information and facilitated mAbs/ACDs candidate selection process
Detailed description:
Characterization of therapeutic monoclonal antibodies (mAbs) or Antibody drug conjugates (ADCs) is challenging due to their ability to bind to a variety of proteins via their Fc and Fab domains, giving rise to diverse biological functions associated with each domain. The Fc domain of mAbs interacts with Fc receptors with varying affinities, which can influence biological processes such as Complement-dependent cytotoxicity (CDC) and Antibody-dependent cellular cytotoxicity (ADCC), transcytosis, phagocytosis, and/or serum half-life.
An important characteristic of an antibody is its Fc effector function. Antibodies can be engineered to obtain desired binding of the Fc region to Fc receptors expressed on effector cells. Hence, it is crucial to evaluate the binding interaction of mAbs/ADC with Fc receptors in the early phase of drug development to understand the potential biological activity of the product in vivo.
Surface Plasmon Resonance (SPR) is a powerful technique to establish binding kinetics in real-time, label free, and high sensitivity with low sample consumption. Along with target antigen binding, it is crucial to evaluate the binding interaction of antibodies and ADCs with Fc receptors. Our SPR case studies investigated the impact on binding kinetics of ADCs with different linkers and the binding interactions of SARS-CoV-2 spike protein variants and evaluated the neutralizing ability of therapeutic mAbs. SPR characterisation can be facilitated in all stages of the product life cycle to ensure the quality and safety of mAbs and ADCs.
The Role of BioPhorum Extractables Data in the Effective Adoption of Single-U...Merck Life Sciences
Regulatory expectation does require patient safety evaluations with supporting data for manufacturing components that directly come into contact with drug manufacturing process streams. Readily available extractables data can help manufacturers using singleuse technology to accelerate product qualifications, risk assessments and process optimization
This white paper guides you on how to save time and resources with supplier-provided single-use system extractables data and gives you an overview about the overall strategy for Extractables & Leachables. At the end you will find a case study.
Find more information about filters and single-use components on our website: https://www.sigmaaldrich.com/DE/en/services/product-services/emprove-program/emprove-filter-and-single-use-component-portfolio
Watch the recording of this presentation here: https://bit.ly/3zTOpe4
Detailed description:
SARS-CoV-2 showed us that technology supports us during our inspection activity even if on-site visits are not possible. Travel restrictions of various kinds will remain a risk in the future. The use of new technologies has shown that inspections and audits can be carried out despite these restrictions. We will focus on what possibilities the new technologies offer and take a look at the future of inspections and audits.
In this webinar, you will learn:
• Regulatory overview of remote audits
• The technologies needed to support the audit process
• What types of inspections are possible with the use of these technologies
• How audits may look in the future
Presented by:
Daniel Buescher, Product Manager - Digital Solutions
Moving your Gene Therapy from R&D to IND: How to navigate the Regulatory Land...Merck Life Sciences
Watch the recording of this presentation here: https://bit.ly/3SqOsoP
Novel therapies, including cell and gene therapies, continue to be central to innovation in healthcare and represent the fastest growing area of therapeutic medicine. As a consequence, the number of gene therapies undergoing clinical trials has increased significantly in the last five years.
Manufacturing processes for these novel therapeutics are very complex with a high risk of contamination. Regulatory agencies world-wide have responded by issuing guidance to outline their expectations for development and manufacture of cell and gene therapies. Currently, regulatory guidance is not harmonized globally and can often lead to confusion within industry and increased risk of non-compliance.
In this webinar, we'll answer:
• Which regulatory guidelines do you need to comply for your INDs?
• When do you start implementing GMPs and validated assays?
• How do you get your QC testing strategy ‘right the first time’?
• How do you ensure testing is not your rate limiting step for the IND submission?
Presented by:
Manjula Aysola, Senior Regulatory Consultant
Dr. Alison Armstrong, Sr. Director, Technical and Scientific Solutions
Identity testing by NGS as a means of risk mitigation for viral gene therapiesMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3RijkHC
Detailed description:
Imagine you’ve just completed a manufacturing run for your viral vector. Identity testing is performed to confirm the vector sequence. But when the results come back the data reveals unexpected sequence variants! With an appropriate risk mitigation testing strategy, this situation can be prevented.
The situation described above is not hypothetical, and happens more that you think, costing valuable time and resources.
Investigatory testing has shown that sequence variants present in starting materials (e.g. plasmids) are likely to make their way to the final product. Adequate identification of low-level variants with an appropriately sensitive method is critical in ensuring the quality of the final product. A risk-based testing strategy, in the context of identity, for viral vector manufacturing will be presented, focusing on key testing points. NGS assays for identity and variant detection will be highlighted due to their extremely sensitive nature compared to traditional approaches.
In this webinar, we'll explore:
• Regulatory requirements for identity testing
• NGS applications for identity testing as compared to traditional methods
• A case study on the impact of not establishing a proper risk-based testing strategy
Presented by: Bradley Hasson, Director of Lab Operations for NGS Services
Latest advancements of melt based 3D printing technologies for oral drug deli...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3A2WcH4
The application of polymer excipients in 3D printing manufacturing is usually limited due to the concerns of filament strength, high processing temperature and large scale manufacturing.
Latest technology developments are targeting a direct melt deposition to simplify the process and enable a constant and efficient process. Two different processing approaches will be presented:
The advanced melt drop deposition, where individual three dimensional geometries can be created by depostition of polymer droplets and the MED® 3D printing technology which allows by precise layer-by-layer deposition to produce objects with well-designed geometric structures.
In this webinar, you will learn:
• Latest advancements of melt based 3D printing approaches
• Application examples for the individual technologies
• Deep dive in the MED® 3D printing technology to design dedicated drug release profiles
Presented by:
Dr. Thomas Kipping, Head of Drug Carriers
Dr. Xianghao Zuo, Deputy Director of R&D, Triastek
Viral safety of biologics: What's changing with the ICH Q5A revision?Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3t7X9tg
How does the ICH Q5A revision impact viral safety strategies for biologics?
Biologics continue to grow at a fast pace. Manufactured using cell lines of human or animal origin, these are at risk of viral contamination making safety strategies critical. A comprehensive risk mitigation strategy using multiple orthogonal measures is a regulatory expectation. ICH Q5A, the globally-harmonized guideline outlines the expectations. ICH Q5A is currently being revised to address recent scientific advancements including novel therapeutic modalities, new manufacturing paradigms, updates in viral clearance applications, and alternate detection technologies. We’ll discuss the expected changes and potential impact on viral safety strategies with case studies and examples.
In this webinar, you will learn about:
• The Importance of virus testing in biologics products
• Regulatory landscape, expectations for the Q5A revision
• What's new and changing
• Examples of alternate testing schedules, impact on viral clearance
Presented by:
Manjula Aysola, Senior Regulatory Consultant
Alison Armstrong, PhD, Sr. Director, Technical and Scientific Solutions
Improve Operational Efficiency by Over 30% with Product, Process, & Systems A...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3adaxWh
When implementing new automation systems, organizations must consider things like deployment time, user adoption, and costs.
They must also consider the cost of doing nothing – that is, what competitive advantage is lost in standing still? What time and quality is lost in repetitive, manual tasks rather than an automated, digital workflow? What operational efficiencies are lost?
In this webinar we examine how a product, process, and system agnostic automation platform can be deployed faster than traditional system specific software while bringing greater operational efficiencies (in many cases over 30% improvement).
To remain competitive in the market, biopharma manufacturers must adopt automation and digital technologies, but most plants still have island of automation consisting of independently functioning, standalone unit operations. This results in operational inefficiency, regulatory concerns, and a poor understanding of the process and product life cycle.
Taking the first, right step must include considering risks, costs, timelines, and technology alternatives. Traditional automation approaches tied to specific systems, processes, and products are, by their nature, limited; while an agnostic platform will address current biomanufacturing business challenges and ensure future readiness. With the right platform, a phased automation implementation can yield operational efficiency gains of up to 30% and improved product quality and regulatory compliance.
In this webinar, let's explore:
• Challenges of automation and digital technology adoption
• What a product, process, and system agnostic platform entails
• Applications and benefits of a process orchestration platform
• Ensuring future readiness with process orchestration
Presented by:
Braj Nandan Thakur, Global Product Manager - Automation
Insights from a Global Collaboration Accelerating Vaccine Development with an...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3Nbb5ug
Get insights and best practices from a multinational team establishing a platform for vaccine production. See how a long-term collaboration on a bench-scale process used to produce a Virus Like Particle (VLP) vaccine for SARS-CoV-2 was successfully converted to a robust GMP-compatible, scalable process.
The COVID-19 pandemic further emphasized the need for collaboration in the development of urgently needed vaccines and therapeutics. In this webinar, we take you behind the scenes of our collaboration with Technovax and Innovative Biotech in which a scalable VLP vaccine platform was optimized for use in a production facility in Nigeria in response to the need for local production of SARS-CoV-2 vaccines. The flexibility and robustness of the platform will enable its rapid deployment to support the West African pandemic readiness program. Initial development of the VLP process began in late 2019 and by March 2020, was already adapted for production of a SARS-CoV-2 vaccine.
In this webinar, you will learn:
• About building a priceless collaborative network with integrated solutions
• Virus-Like Particle Vaccines
• Process Development Overview and Challenges
• Pre-clinical Results and Next Steps
Presented by:
Jose M. Galarza, PhD,
President and Founder of TechnoVax
Naomi Baer,
Business development consultant, Emerging Biotech, BioProcess division
Youssef Gaabouri, Eng. ,
Associate Director, Head of Sales Middle East & Africa, BioProcess division
Risk-Based Qualification of X-Ray Sterilization for Single-Use SystemsMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3vQf0qv
In the single-use bioprocess industry, X-ray irradiation warrants consideration as an alternate sterilization technology. Using a risk-based qualification testing strategy is important when evaluating and implementing equivalent ionizing irradiation sterilization methods.
The urgent need for life-saving therapies as a result of the global pandemic has reinforced the criticality of flexibility in pharmaceutical manufacturing, including sterilization. The single-use bioprocess industry traditionally has employed gamma irradiation sterilization. X-ray irradiation is being considered as an additional sterilization technology for business and supply continuity. We will share a risk-based qualification testing strategy including Extractables and data generated to support comparability of gamma irradiation and X-ray irradiation as equivalent ionizing irradiation sterilization methods.
In this webinar, you will learn about:
• The comparison of gamma and X-ray irradiation sterilization
• A risk-based qualification test strategy
• Data evaluation of gamma versus X-ray sterilized single-use components
Presented by:
Monica Cardona,
Global Senior Program Manager
Paul Killian, Ph.D.,
R&D Director, Analytical Technologies
Rapid replication competent adenovirus (rRCA) detection: Accelerate your lot ...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3MJ4u9V
Testing for presence of replication competent adenovirus (RCA) is a key component to ensure patient safety and a requirement for all biologicals manufactured using adenoviral vectors. For many adenoviral-based products, the RCA assay is a rate-limiting assay for lot release.
Join this webinar to learn about a rapid RCA detection assay currently in development, which combines a 7-day culture assay with a highly sensitive molecular endpoint specific for RCA. The method can detect presence of as little as 1 RCA in adenoviral vector material at an approximate concentration of 5x107 - 2x108 vector particles (VP)/mL, making it a suitable method to meet regulatory requirements while accelerating your lot release timelines.
In this webinar, you will learn about:
• Regulatory framework for adenoviral vector products
• Considerations for lot release testing of adenoviral-based therapies
• Advantages of a rapid method for RCA testing on production lot material
Presented by:
Axel Fun, Ph.D.,
Principal Scientist
Alberto Santana, MBA,
Product Manager, Biologics Biosafety Testing
The High Intensity Sweeteners Neotame and Sucralose: 2 Ways to ace the Patien...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3vQyN7K
Bitter medicines are an important issue, especially for pediatric applications. As several APIs have bitter tasting components, high intensity sweeteners for taste optimization are of great interest. Join our webinar to discover our new sweetener toolbox enabling safe and stable formulations.
Mask bitter aftertaste for a sweeter pill to swallow! Patients’ compliance and the therapeutic benefit are supported by a pleasant taste of pharmaceutical formulations. With the high intensity sweeteners Neotame and Sucralose, you have efficient tools at hand which are superior to other sweeteners in many aspects:
• excellent sugar-like taste profile
• outstanding sweetness factors
• use effectiveness
• enhanced stability
We will present our new toolbox of two high performance sweeteners and focus on aspects of stability, safety, the application in various dosage forms, and market perception.
In this webinar, you will learn:
• How to optimize the patients' taste experience of your pharmaceuticals
• How sweeteners can be differentiated by their sensory profiles and features
• How our new product offering Neotame can be effectively used in your targeted formulations
Presented by:
Almut von der Brelie,
Senior Manager Strategic Marketing
Excipients for Solid Applications
The Developability Classification System (DCS): Enabling an Optimized Approac...Merck Life Sciences
This whitepaper by Dr. Daniel Joseph Price outlines how poorly soluble drug formulations can be designed using the developability classification system (DCS).
The DCS identifies the root cause of low solubility and enables lean, cost-effective and effective formulations to be developed.
#solubility #pharmaceuticalmanufacturing #oralsoliddosage #drugdevelopment
In this webinar, you will learn about:
The advantages of using advanced intermediates to develop ADC therapies
How to increase ADC solubility and efficiency
Fast, small-scale ADC library generation
Seamless supply chain with reduced complexity and regulatory support
The ADCore product line offers versatile intermediates that simplify the synthesis of common ADC payloads (dolastatins, maytansinoids, and PBDs) by greatly reducing the number of synthetic steps. This translates to savings in development and manufacturing costs and shorter timelines to the clinic. To address the poor solubility of many ADC payloads, ChetoSensar™ was developed to significantly increase the hydrophilicity of the drug linker, which has been shown to also substantially increase the efficacy of ADCs and broaden the therapeutic window.
Lastly, the ADC Express™ service leverages conjugation chemistry and analytical expertise to help design and quickly synthesize sets of potential ADC therapies suitable for screening to simplify candidate selection and get ADC therapies to market faster.
Regulatory Considerations for Excipients used in Lipid NanoparticlesMerck Life Sciences
Lipid excipients and delivery systems such as lipid nanoparticles (LNPs) are essential for a wide variety of therapeutics including mRNA vaccines and therapeutics and gene therapy.
The purity and safety of novel, synthetic lipid excipients must be demonstrated due to their central role in the function of the drug product, distinct physicochemical properties, and the potential for interaction with other ingredients or the physicochemical environment. These excipients must comply with challenging and complex regulatory requirements, similar to those expected of the active pharmaceutical ingredient itself.
This whitepaper provides an overview of the regulatory classification of lipid nanoparticles, liposomes and novel excipients. Specific requirements outlined in guidance documents are shared along with strategies to stay ahead of emerging regulatory challenges.
To find more information about synthetic lipids for pharmaceutical applications and gene therapy, please visit our website:
https://www.sigmaaldrich.com/DE/en/products/pharma-and-biopharma-manufacturing/formulation/synthetic-lipids
https://www.sigmaaldrich.com/US/en/products/pharma-and-biopharma-manufacturing/formulation/synthetic-lipids
Defecation
Normal defecation begins with movement in the left colon, moving stool toward the anus. When stool reaches the rectum, the distention causes relaxation of the internal sphincter and an awareness of the need to defecate. At the time of defecation, the external sphincter relaxes, and abdominal muscles contract, increasing intrarectal pressure and forcing the stool out
The Valsalva maneuver exerts pressure to expel faeces through a voluntary contraction of the abdominal muscles while maintaining forced expiration against a closed airway. Patients with cardiovascular disease, glaucoma, increased intracranial pressure, or a new surgical wound are at greater risk for cardiac dysrhythmias and elevated blood pressure with the Valsalva maneuver and need to avoid straining to pass the stool.
Normal defecation is painless, resulting in passage of soft, formed stool
CONSTIPATION
Constipation is a symptom, not a disease. Improper diet, reduced fluid intake, lack of exercise, and certain medications can cause constipation. For example, patients receiving opiates for pain after surgery often require a stool softener or laxative to prevent constipation. The signs of constipation include infrequent bowel movements (less than every 3 days), difficulty passing stools, excessive straining, inability to defecate at will, and hard feaces
IMPACTION
Fecal impaction results from unrelieved constipation. It is a collection of hardened feces wedged in the rectum that a person cannot expel. In cases of severe impaction the mass extends up into the sigmoid colon.
DIARRHEA
Diarrhea is an increase in the number of stools and the passage of liquid, unformed feces. It is associated with disorders affecting digestion, absorption, and secretion in the GI tract. Intestinal contents pass through the small and large intestine too quickly to allow for the usual absorption of fluid and nutrients. Irritation within the colon results in increased mucus secretion. As a result, feces become watery, and the patient is unable to control the urge to defecate. Normally an anal bag is safe and effective in long-term treatment of patients with fecal incontinence at home, in hospice, or in the hospital. Fecal incontinence is expensive and a potentially dangerous condition in terms of contamination and risk of skin ulceration
HEMORRHOIDS
Hemorrhoids are dilated, engorged veins in the lining of the rectum. They are either external or internal.
FLATULENCE
As gas accumulates in the lumen of the intestines, the bowel wall stretches and distends (flatulence). It is a common cause of abdominal fullness, pain, and cramping. Normally intestinal gas escapes through the mouth (belching) or the anus (passing of flatus)
FECAL INCONTINENCE
Fecal incontinence is the inability to control passage of feces and gas from the anus. Incontinence harms a patient’s body image
PREPARATION AND GIVING OF LAXATIVESACCORDING TO POTTER AND PERRY,
An enema is the instillation of a solution into the rectum and sig
CRISPR-Cas9, a revolutionary gene-editing tool, holds immense potential to reshape medicine, agriculture, and our understanding of life. But like any powerful tool, it comes with ethical considerations.
Unveiling CRISPR: This naturally occurring bacterial defense system (crRNA & Cas9 protein) fights viruses. Scientists repurposed it for precise gene editing (correction, deletion, insertion) by targeting specific DNA sequences.
The Promise: CRISPR offers exciting possibilities:
Gene Therapy: Correcting genetic diseases like cystic fibrosis.
Agriculture: Engineering crops resistant to pests and harsh environments.
Research: Studying gene function to unlock new knowledge.
The Peril: Ethical concerns demand attention:
Off-target Effects: Unintended DNA edits can have unforeseen consequences.
Eugenics: Misusing CRISPR for designer babies raises social and ethical questions.
Equity: High costs could limit access to this potentially life-saving technology.
The Path Forward: Responsible development is crucial:
International Collaboration: Clear guidelines are needed for research and human trials.
Public Education: Open discussions ensure informed decisions about CRISPR.
Prioritize Safety and Ethics: Safety and ethical principles must be paramount.
CRISPR offers a powerful tool for a better future, but responsible development and addressing ethical concerns are essential. By prioritizing safety, fostering open dialogue, and ensuring equitable access, we can harness CRISPR's power for the benefit of all. (2998 characters)
Navigating Challenges: Mental Health, Legislation, and the Prison System in B...Guillermo Rivera
This conference will delve into the intricate intersections between mental health, legal frameworks, and the prison system in Bolivia. It aims to provide a comprehensive overview of the current challenges faced by mental health professionals working within the legislative and correctional landscapes. Topics of discussion will include the prevalence and impact of mental health issues among the incarcerated population, the effectiveness of existing mental health policies and legislation, and potential reforms to enhance the mental health support system within prisons.
R3 Stem Cells and Kidney Repair A New Horizon in Nephrology.pptxR3 Stem Cell
R3 Stem Cells and Kidney Repair: A New Horizon in Nephrology" explores groundbreaking advancements in the use of R3 stem cells for kidney disease treatment. This insightful piece delves into the potential of these cells to regenerate damaged kidney tissue, offering new hope for patients and reshaping the future of nephrology.
Medical Technology Tackles New Health Care Demand - Research Report - March 2...pchutichetpong
M Capital Group (“MCG”) predicts that with, against, despite, and even without the global pandemic, the medical technology (MedTech) industry shows signs of continuous healthy growth, driven by smaller, faster, and cheaper devices, growing demand for home-based applications, technological innovation, strategic acquisitions, investments, and SPAC listings. MCG predicts that this should reflects itself in annual growth of over 6%, well beyond 2028.
According to Chris Mouchabhani, Managing Partner at M Capital Group, “Despite all economic scenarios that one may consider, beyond overall economic shocks, medical technology should remain one of the most promising and robust sectors over the short to medium term and well beyond 2028.”
There is a movement towards home-based care for the elderly, next generation scanning and MRI devices, wearable technology, artificial intelligence incorporation, and online connectivity. Experts also see a focus on predictive, preventive, personalized, participatory, and precision medicine, with rising levels of integration of home care and technological innovation.
The average cost of treatment has been rising across the board, creating additional financial burdens to governments, healthcare providers and insurance companies. According to MCG, cost-per-inpatient-stay in the United States alone rose on average annually by over 13% between 2014 to 2021, leading MedTech to focus research efforts on optimized medical equipment at lower price points, whilst emphasizing portability and ease of use. Namely, 46% of the 1,008 medical technology companies in the 2021 MedTech Innovator (“MTI”) database are focusing on prevention, wellness, detection, or diagnosis, signaling a clear push for preventive care to also tackle costs.
In addition, there has also been a lasting impact on consumer and medical demand for home care, supported by the pandemic. Lockdowns, closure of care facilities, and healthcare systems subjected to capacity pressure, accelerated demand away from traditional inpatient care. Now, outpatient care solutions are driving industry production, with nearly 70% of recent diagnostics start-up companies producing products in areas such as ambulatory clinics, at-home care, and self-administered diagnostics.
Nursing Care of Client With Acute And Chronic Renal Failure.ppt
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing
1. The life science business of Merck KGaA,
Darmstadt, Germany operates as
MilliporeSigma in the U.S. and Canada.
Data driven strategies and
considerations for scalable
purification of Plasmid DNA
for use in vaccine
manufacturing
Thomas Parker, Global Focal Point – Vaccines and Viral Therapies,
Technology Management
29 October 2020
2. The life science business
of Merck KGaA, Darmstadt,
Germany operates as
MilliporeSigma in the U.S.
and Canada
5. • Circular double helix DNA molecules, naturally found in bacteria, intracellular replicated
• Molecular characteristics:
− Large size 1.5 – 150 kb
− Poly anion, highly negatively charged
− Sensitive to mechanical stress
− Various topological forms
ccc form I: covalentely closed circles, supercoiled, fully intact,
wound around itself
oc form: one strand nicked/broken, less compact, totally relaxed
linear form: both strands broken, free ends
Introduction
What are Plasmids?
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing5
MW Size
Plasmid 4 MD* 200 nm**
mAB 0.2 MD* 10 nm**
* Mega Dalton (6 kb Plasmid)
** Dynamic range
Supercoiled plasmid is recognized by FDA as the most therapeutically effective
Carsten Voß, 2007
6. Importance of Plasmid DNA
The key-role of pDNA for viral vector and vaccine production
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing6
Plasmid DNA
(Vector)
Therapeutic
transgene, structural
genes
CAGR 18%
Other
14%
Adeno
15%
LV
49%
pDNA
22%
Viral Vectors
Clinical trials by vector types
Bioprocess International,
December 17, 2019
DNA vaccines
▪ - prophylactic use,
e.g. infections
- therapeutic
applications,
e.g. cancer, allergy
mRNA vaccines
7. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing7
✓ Capable of inducing strong
immune response
✓ Require appropriate formulation
(buffers, stabilizers, and inorganic or
organic matrices) for protection
from degradation and enable
efficient transfer and delivery
Plasmid DNA as a Vaccine
Basic Mechanism
Delivery methods: lipofection, gene-gun (DNA coated gold particle,
micro-particles/capsules)
1) Viral protein
gene transferred
into a plasmid
2) Plasmid purified
and delivered to
patient skin or
muscle cells
3) Patient cells produce
the viral protein,
inducing an immune
response
8. Advantages
Challenges
Downsides
Characteristics
Plasmid DNA as a Vaccine
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing8
• Simple design
• Easy to manipulate/optimize
• Production: Reproducible, rapid,
scalable
• Requires only BL1 safety level
• No significant adverse events
• Temperature stable, long shelf life
• Similar immune response as other
platforms
• Removal of impurities and non
desired pDNA forms need careful
process design
• High concentration needed (mg
level)
• Delivery and protection of DNA
from degradation
• Poor gene transfer efficiency
• Estimated that for every 1000
plasmid molecules only one reaches
the cell and is expressed.
• Low immunogenic potential of
bacterial DNA
• Possible insertional mutagenesis
9. Importance of Plasmid DNA Vaccines
Fighting COVID 19
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing9
Four pDNA based vaccines in clinical evaluation
12 pDNA vaccines in preclinical evaluation
WHO report, 02 October 2020
COVID-19 Vaccine
developer/manufacturer
Type of candidate
vaccine
Number of
doses
Timing of
doses
Route of
Administration
Clinical Stage
Inovio Pharmaceuticals/
International Vaccine Institute
DNA plasmid vaccine with
electroporation
2 0, 28 days Intradermal I/II
Osaka University/ AnGes/ Takara
Bio
DNA plasmid vaccine +
Adjuvant
2 0, 14 days intramuscular I/II
Cadila Healthcare Limited DNA plasmid vaccine 3 0, 28, 56 days Intradermal I/II
Genexine Consortium
DNA Vaccine
(GX-19)
2 0, 28 days Intramuscular I/II
10. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing10
Plasmid Production
Unique challenges of pDNA purification
Adequate technologies and application expertise for providing solutions for Plasmid
manufacturing needed.
• Low productivity of microbial fermentation due to copy number limitations in the relevant strains.
• Similarity of product and contaminants (gDNA, Endotoxin, RNA, Plasmid isoforms) leads to low
resolution separation.
• Feed often highly viscous, complicating downstream processing.
• Low flowrate needed for chromatography
• Difficult to achieve desired concentrations at the final TFF step.
• Shear sensitivity.
• Need to eliminate multiple undesirable components from the process.
• Lack of platform process and integrated solutions.
From: Xenopoulos, A. and Pattnaik, P. (2014), Expert Rev. Vaccines 13, 1537-1551
11. Unit Operation BI LONZA Cobra bio Merck & Co. Inovio GSK GEHC BIA
Plasmid pRZ-hMCP1 pTX0161 pVAX1 Pegfp-N1
Host K12 JM108 Clean Genome DH1 DH1 OB DH1 DH5a
Cell Harvest Continuous
centrifugation
centrifugation centrifugation TFF (0.45 UM)
Cell lysis Continuous alkaline
lysis
Continuous alkaline
lysis
Alkaline lysis with
RNase
Heat lysis with
Lysozyme
Continuous alkaline
lysis
Alkaline lysis Alkaline lysis Alkaline lysis
Primary Clarification Flocculation Woven nylon bag filter Bubble column with
3/0.1 um depth filters
Centrifugation>membr
ane (0.45/0.2 um)
concentration Ultrafiltration (hollow
fiber)
TFF Ultrafiltration (hollow
fiber, 100 kD)
Contaminant
Precipitation
Ammonium sulphate Chemical CTAB Fractional Calcium chloride Calcium chloride Calcium chloride
Sec.Clarification filtration Depth filter Filtration with DE Depth filtration
1.2/0.45/0.2 um
membranes
Concentration Hollow fiber TFF TFF 100 KD
Chromatography HIC-Butyl 650 M, B/E.
AEX: Fractogel DEAE
B/E
SEC-Sepharose 6FF
HIC-hexyl 650 C F/T.
HIC-Butyl 650 S, B/E.
SEC-Sepharose 6FF
AEX; DEAE Expanded
bed
SEC S500HR
AEX : Q Membrane,
B/E
HIC: Resin B/E
AEX: Fractogel TMAE SEC: Sepharose 6FF
media
Thiophilic aromatic
plasmid Select B/E
AEX: Source 3 0Q, B/E
AEX: DEAE Monolith
HIC: monolith
Concentration TFF TFF (Spiral device, 20
kD)
Ethanol Precipitation
and TFF,RC, 100 Kda
TFF-PES, 50 kD
Final filtration PVDF 0.22 PVDF 0.22 PES
No industry platform solution has been accepted
Diversity of existing process schemes
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing
11
Large-Scale Production Processes for Plasmid DNA
From: Xenopoulos, A. and Pattnaik, P. (2014), Expert Rev. Vaccines 13, 1537-1551
12. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing12
Generic Production Process for Plasmid DNA
Plasmid Overview
Fermentation Harvest Lysis
Alkaline Lysis
or
Heat,
Mechanical,
Enzymatic
Clarification/Primary recovery
Purification
Capture, Polishing
Sterile Filtration
Centrifugation
Filtration
Flocculation, ATPS,
Selective Precipitation (PEG, salt
detergent), Expanded Bed
Optional Feed pre-treatment
Centrifugation
TFF or
hollow fiber
TFF
Concentration,
Diafiltration
AEX -> HIC
HIC -> AEX
SEC -> HIC
Affinity
1-2 Chromatography steps
By
CaCl2-Preciptitation
or RNAse
Concentration Buffer
exchange by TFF
Adjustment
for Capture
RNA removal
13. Integrated strength from harvest to final fill
Our Capabilities for pDNA Purification
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing13
Fermentation Clarification
Purification with
Chromatography(1-2 steps)
Concentration and
Diafiltration (UF/DF)
Final filtrationStorage Concentration and
diafiltration (UF/DF)
Thaw cells Cell harvest Cell lysis
Prostak®/Pellicon® Mobius® Mixer
Clarisolve®/Millistak+®
Pellicon® 2 100 or 300 kDaFractogel® DEAE
Fractogel® DMAE
Natrix® Q
Pellicon® 2 100 kDaMillipore Express®
Validation and Testing Services Mobius® Single Use Solutions
14. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing14
Poll Question #1
16. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing16
Plasmids are produced
intracellularly
• Multiple unit operations are needed before
downstream purification
Cells harvest by
centrifugation or
tangential follow
filtration (TFF)
microfiltration
Cell lysis by
physical or
chemical
treatment
Clarification by
depth filtrating
or centrifugation
Harvest overview
Cell Harvest, Lysis, & Clarification
17. • Plasmids are produced intracellularly in a batch and/or
fed-batch using E. coli host
• Minimal mineral salt media commonly used - low price,
autoclavable
• Productivity and specific productivity (amount of pDNA per
unit of cell mass) are proportional to the final cell density in
a fermenter
• Fast production (~1 day) relative to mammalian cell
culture
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing17
Fermentation overview and output
Cell Harvest, Lysis, & Clarification
Defining the harvest input:
• Wide range of volume possible
• 30-200L common, but
6000L proven
• OD600nm of 100-120
• 100-250 mg/L, yields up to
2 g/L reported
• Specific yield of 50 mg
pDNA/g DCW, up to 75% of
total DNA is Plasmid
18. Microfiltration Tangential Flow Filtration
(MF-TFF)
High solids count favor open-channel formats
Flat-sheet TFF devices work well in this
application
− Linear scalability
− Wide range of formats and installation options
Centrifugation
Traditional approach is cost effective at large
scale (>500L) or very small scale with swinging
bucket (<5L)
Special attention is needed due to high shear
generation in large scale centrifugation
Centrifugation vs Tangential Flow Filtration
Cell Harvest, Lysis, & Clarification
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing18
Parameters Value
Device
Prostak™ or Pellicon® 2 with Durapore®
0.1, 0.22 µm, or 0.45 µm
Pellicon® 2 Biomax® 1000 kDa, V screen
Pellicon® 2 Ultracel® 1000 kDa, V screen
Volumetric loading 10 – 60 L/m2
Feed flow 7 - 9 L/min/m2
TMP < 0.5 bar
Average flux 20 - 30 LMH
Volumetric
concentration factor
2 to 5
Diafiltration volumes 3 to 5
MF-TFF Typical Operating Parameters
19. Principle
• Alkaline condition plus detergent solubilizes the cell walls and
the alkaline environment denatures genomic DNA.
• Sodium dodecyl sulfate and Triton X-100 common
• Reaction neutralized with acid after short incubation
Challenges
• Both alkaline and shear from mixing will damage supercoiled
plasmid DNA
• Sudden increase of viscosity will impact mixing efficiency
Solutions
• Optimize mix speed, incubation time, and chemical
concentrations
• Mobius® single use mixers
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing19
Key considerations for lysis
Cell Harvest, Lysis, & Clarification
• Lysis affects pDNA quality and
impurity level, directly
impacting subsequent down
stream steps.
• Continuous lysis approach
may be needed for large
scales
20. Aims at reducing lysate impurity level for improving
clarification and purification by use of:
• PEG/PEI for selective precipitation of gDNA
• Divalent cations (2M Ca2+) for precipitation of RNA
• RNAse for enzymatic digestion RNA (cost and regulatory
concerns)
• Anionic detergent for lowering endotoxin levels
Watch Outs
• Changes of lysate pH, salt conc. conductivity, viscosity
• Additives might not be compatible with subsequent
chromatography capture and require additional TFF for
buffer exchange
• Additives might be difficult to remove
Cell Harvest, Lysis & Clarification
Optional Pretreatment
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing20
Pretreatment impacts clarification performance and can affect
subsequent chromatography capture step.
Ca+2 mediated
RNA precipitation
AEX Capture
Clarification
AEX Capture
vs.
LysateLysate
TFF
Clarification
21. • Lysis output can be challenging for clarification
• Complex mixture:
• Large particles, cell debris, plasmid DNA,
genomic DNA, RNA and host cell proteins
• Very high solids load
• Highly viscous
• Two-phase separation
• Froth phase (top) – cell debris and gDNA
• Lysate phase (bottom) – pDNA, RNA, HCP
• Manufacturers utilize a wide range of pre-
clarification methods to enhance unit operation
efficiency
Cell Harvest, Lysis & Clarification
No platform solution for lysis and conditioning
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing21
Internal Data
25%
10%
10%
15%
25%
15%
Pre-clarification conditioning
Sieving/Filtration Centrifugation
Polymer Flocculation Natural Floating Separation
No Pre-clarification Proprietary
22. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing22
Filtration based
clarification
preferred up to
500L+
fermentation
scale Modern approach, use
size exclusion and
absorption mechanisms
Can be used in series
Attention: positive
charged filtration aid
may interact with
plasmid
Centrifugation vs Filtration
Cell Harvest, Lysis & Clarification
Traditional approach,
which is capable of
handling high %solids
May require secondary
clarification
Attention: high shear
rate can damage the
structure of supercoiled
plasmid DNA
Centrifugation Normal flow filtration
23. Depth filters at a glance
Cell Harvest, Lysis & Clarification
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing23
24. Depth filters at a glance
Cell Harvest, Lysis & Clarification
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing24
Media type and density impact adsorptive properties
Lower adsorption Higher adsorption
25. Capacity range for non-conditioned lysateCapacity range for pre-conditioned lysate
Data summary for clarification using depth filtration
Cell Harvest, Lysis & Clarification
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing25
0
50
100
150
200
250
300
350
Clarisolve
20MS
Clarisolve
40MS
Clarisolve 60
HX
Millistak+
D0HC
Millistak+
C0HC
Capacity(L/sqm)
0
50
100
150
200
250
300
350
400
450
Clarisolve
40MS
Clarisolve
60HX
Millistak+
D0HC
Millistak+
C0HC
Millistak+
D0SP
Millistak+
CE50
Capacity(L/sqm)
High yields at >90%
• Millistak+® D0HC or C0HC: >80%
• Millistak+® CE: >90%
• Clarisolve®: >90%
• Salt chase recommended
• Bioburden reduction
membrane capacity post
depth filtration varies –
>500L/m2 typical
26. Case Study #1
Clarisolve® 60HX
Alkaline lysis + CaCl2 precipitation
− gDNA and RNA being removed
Case Study #2
Millistak+® D0HC
Alkaline lysis without pretreatment
− gDNA removed
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing26
For 500L, Case #1 needs two
Process Scale Pod Racks
and Case #2 needs one
Case studies
Cell Harvest, Lysis & Clarification
Filter
Flux
(LMH)
Volumetric
Throughput at 15psi
dP (L/m2)
Installation for
500L with 1.5x
safety factor
Case #1 Clarisolve® 60HX 200 160 14 x 0.55m2
Case #2 Millistak+® D0HC 100 150 8 x 1.1m2
28. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing28
Poll Question #2
29. General Considerations
Plasmid Purification
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing29
Polish
Chromatograph
HIC
AEX
Ultrafiltration/
Diafiltration
Final filtrationFill in
assemblies
Optional
Ultrafiltration
Capture
Chromatography
AEX
HIC
Plasmid purification requires a combination of more
than one unit operation.
Tangential Flow Filtration (TFF) at 1-2 steps
• Can remove RNA, small sized genomic DNA and protein
• Essential for concentration to reduce loading time and
complete buffer exchange prior chromatography steps
• After clarification and before chromatography
Chromatography
• Selectively separates supercoiled Plasmid from oc-/linear
isoforms and residual impurities (host cell protein and nucleic
acid, endotoxin) by charge, size or hydrophobicity
• Anion Exchange (AEX) and Hydrophobic interaction (HIC)
typically used
• Sometimes size exclusion (SEC) for RNA removal
Sterile filtration to ensure process safety
or
Biggest Challenge:
Separation of Plasmid isoforms
31. Ultrafiltration and Diafiltration Overview
TFF
30, 100 or 300 kD membranes can be used at two different steps
First TFF step, after clarification and before chromatography
− Remove residual impurities and protein
− Concentrate plasmid to reduce loading time and buffer exchange for
chromatography steps
− 4-25x concentration and 5-10 diavolumes
Second TFF step, after chromatography and before sterile filtration of
drug substance
− Achieve final concentration and exchange into formulation buffer
− 10x concentration and ~10 diavolumes typical
31
1
2
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing
32. Two-pump control adds operational consistency
TFF
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing32
Use 2-pump control for 100kD
and higher
When low TMP causes Critical Permeate
Flux, 2-pump control is recommended
TMP control has difficulties maintaining a stable
system at low TMP (~5psi or less)
Very tight window of operation below ~5psi TMP
Deviation from TMP setpoint could result in permeate flux
near or above critical flux
2-pump control allows consistent operation
Ensures permeate flux does not approach
critical flux
Minimize fouling
Consistent impurity clearance and yield
0
2
4
6
8
10
12
14
30kD 100kD 300kD 1000kD
TMP(psi)
TMP @ Critical Flux
4LMM 5.5LMM 7LMM
33. Parameter Value
Feed Flux (LMM) 4 or less
Permeate Flux (LMH) – for
two pump
20-50
TMP (psi) – for TFF control <10
Loading (L/m2) 70-140
Concentration Factor 5x-50x
TFF
Recommended operating parameters
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing33
Watch out for:
• High viscosity
• Shear
D.R. Latulippe, A.L. Zydney / Journal of Colloid and Interface Science 357 (2011) 548–553
Operating parameters can
change retention properties:
35. Common Approaches at Large Scale
Chromatography
35
Anion Exchange Chromatography (AEX)
• Well suited for binding polyanionic DNA
• Applicable for capture, intermediate and polishing
• Weak AEX resins (DEAE, DMAE) give highest recovery and selective impurity removal
o Resins with higher ionic capacity can show recovery issues
• Charge similarity of Plasmid and impurities poses a challenge
• Capable of separating plasmid from proteins, RNA and gDNA and removing Endotoxin
• Separation of Plasmid isoforms difficult
Hydrophobic Interaction Chromatography (HIC)
• Works by salt promoted binding (≈ 2.5 M NH4SO4)
• Supercoiled pDNA is less hydrophobic than RNA, oc- and linear- Plasmid forms and denatured gDNA
• Conveniently used for intermediate purification after AEX capture due to high salt AEX eluate pool
o Can also be used for primary capture
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing
36. Membrane Chromatography
• Improved hydraulic performance due to large
convective pores and low bed height
• Large accessible surface for Plasmids enabling for
improved binding capacity (5 - 10 mg/mL)
at very short residence times < 0.2 min
• High productivity due to short cycling time
• Single use format, principally scalable
Two technologies to consider
Chromatography
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing36
Resin Chromatography
• Low binding capacity due to large Plasmid size
limiting diffusion into beads restricting binding to the
resin surface only
• High pressure drops due to elevated viscosity feed
stream resulting in flow limitations and long
processing times (2-8min RT typical)
• Re-use needed for economic feasibility
• Flexible installation – pack as much as needed
• Generally better selectivity than membranes
37. Solutions
Chromatography Products for Plasmid Purification
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing37
Single-use solution providing superior
productivity and flexibility to meet the
challenges of multi-product / multi-purpose
manufacturing of Plasmids
Ideally suited for the capture of small and
mid-sized Plasmids (<20kbp)
Versatile solution offering unique
selectivity and best in class capacity
Efficient and cost-effective operation,
however, require optimal column sizing
(in order to accomplish rapid cycling and
realize low hardware costs)
Fractogel® DEAE/DMAE (wAEX)Natrix® Q
38. • DMAE and DEAE ligands exhibit unique binding
selectivity
• Moderate binding capacity of 2 – 4 mg/mL
• High yield: 80% to 95% of ccc-form
• Removal of residual Endotoxin
• Good resolution due to moderate bead size
(d50: 48 - 60 µm)
• Moderate flow: 4-8 min residence time
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing38
✓ High surface area of tentacle resin
offer improved capacity relative to
conventional anion exchange resins
due to tentacles
✓ Proven resins for reliable Plasmid
purification
Plasmid Purification
Fractogel® DEAE and DMAE Chromatography Resin
Ideal for intermediate purification / polishing
but still frequently used for capture.
39. Optimal amount of salt supplement:
• depends on resin type
• varies with initial lysate conditions
(protocol specific lysis buffer)
• gives higher binding capacity and
improved Plasmid eluate purity
• should be determined for each specific
lysate condition by screening
Optimal Conditions for direct AEX Capture
Fractogel® DEAE and DMAE Chromatography Resin
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing39
0.0
1.0
2.0
3.0
4.0
5.0
0 100 200 300 400 500
PlasmidSBC(mg/mL)
NaCl supplement (mM)
FG DEAE (M)
FG DMAE (M)
1 Marker
2 0 mM NaCl
3 100 mM NaCl
4 120 mM NaCl
5 150 mM NaCl
Purity of Fractogel® DEAE
capture eluates from lysates
with varying NaCl supplement
Binding of Plasmid
from lysate
Salt supplementation of lysate is a simple and efficient method to prevent RNA
binding and reduce interference with plasmid binding on AEX
40. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing40
Commercial Plasmid Production
Preferred Chromatography Approach
• Lysate with 120 mM NaCL
supplement
• Capacity of ~2.5 mg Plasmid /
mL CV
• > 95% RNA in flow through
• > 95% pDNA purity in eluate
• > 90% pDNA yield by salt
step gradient
• Pre-elution wash step including
alcohols (e.g. 2-Propanol)
required for improved endotoxin
removal
AEX Capture
Fractogel® DEAE or
Fractogel ® DMAE
Removes RNA, endotoxin,
gDNA, host cell proteinSupplementation
of NaCl
HIC Polishing
Separates Plasmid
isoforms
Clarified lysate
pH 5.0, 67 mS/cm,
1M K-acetate
adjust to 2.4M
NH4SO4
Plasmid Capture with Fractogel® DEAE (M)
Salt supplementation for AEX direct capture
eliminates need for pre-treatment and TFF
41. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing41
Alternative Chromatography Approach
Commercial Plasmid Production
pDNA binding to Fractogel® AEX resins
tolerates presence of elevated
concentration of (NH4)2SO4
Precipitate formation upon lysate
adjustment to HIC conditions is an issue,
involves additional effort for clarification
AEX Polishing
Fractogel® DEAE or Fractogel ® DMAE
Removes residual RNA, endotoxin, gDNA, host cell
protein
HIC Capture
Clarified lysate
pH 5.0, 67 mS/cm, 1M K-acetate
Adjustment to HIC binding conditions
*Data using purified pDNA, 4.7kbp
42. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing42
Natrix® Q Chromatography Membrane in Single-Use Format
High-throughput Capture
Plasmid Application
• High capacity of 5-10 mg/mL at only 0.1 min
residence time possible with small (5.7 kb) and
mid-sized Plasmids (13kb)
• Yield: ≥80 % of ccc-form
• > 95 % RNA removal
• Short cycle time: 35 min
• Caustic stability enables for efficient cleaning
(1M NaOH +2M NaCl)
• Re-use in rapid-cycling operation mode allows for
cost reduction
• Narrow pore size poses challenge for larger Plasmids
Material Properties
• Composite membrane
• High mechanical strength and hydraulic
permeability
• Nominal pore size: 0.4 µm
• Permits high flow
• Recommended 10MV/min,
(6 sec. residence), range of 5-25 MV/min
43. Performance Overview - Examples
Plasmid Purification with Natrix® Q Chromatography Membrane
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing43
Feed used:
• Original E. coli lysates from alkaline lysis, clarified by centrifugation/depth filtration, supplemented with optimal NaCl
to eliminate RNA interference, pH 5.0, finally 0.22 µm filtered
• Plasmids ranging from small to large size
• Varying initial pDNA purity ranging from 0.7% - 4%
For Plasmids with 20 kb size or larger, a collapse of membrane permeability was encountered with loadings greater
than 2 mg/mL.
Plasmid
Size
Plasmid
Titer
(µg/mL)
Initial pDNA
Purity
(A260 based)
Initial Lysate
Conductivity
(mS/cm)
Optimal NaCl
Supplement
(mM)
Final Lysate
Conductivity
(mS/cm)
Residence
Time
(min)
Operating
Capacity
(mg/mL MV)
Yield
(%)
pDNA Eluate Purity
(A260 based)
5.7 kb 45 4.0 % 69 160 82 0.1 10 ≥ 80 ≥ 80 % pDNA
13 kb 33 2.4 % 72 130 82 0.1 4 ≥ 77 ≥ 90 % pDNA
20 kb 25 0.7 % 79 100 86 0.2 1 ≥ 65 ≥ 62 % pDNA
44. Performance Overview - Examples
Chromatography Products for Plasmid Purification
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing44
Recommended
Process Step
Resin /
Membrane
Adsorber
Dynamic
Capacity
(mg/mL)
Cycle
Time
(min)
Residence
Time
(min)
RNA
Remov
al
Yield
ccc-
Form
Purity
(A260
based)
High-
throughput
capture
Natrix® Q 10 0.5 h 0.1 – 0.03 >95 % ≥80 %
>80 %
pDNA
Polishing
Purification
Fractogel®
DEAE (M)
2.5 3 h 4 >95 % ≥80 %
>95 %
pDNA
Fractogel®
DMAE (M)
3 3 h 4 >95 % ≥95 %
>95 %
pDNA
Feed used:
Original E. coli lysate, clarified by centrifugation and subsequent depth filtration, directly
supplemented with NaCl (120 – 250 mM, depending on resin or membrane type) to eliminate RNA
interference, pH 5.0, 74 – 82 mS/cm
pDNA size 5.7 kbp, pDNA titer 45 µg/mL
* 10 cm bed height
46. Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing46
Sterile Filtration
Overview of pDNA Sterile Filtration
Recommendations
✓Millipore Express® SHC (PES)
over Durapore® 0.22um (PVDF)
✓Optimize conditions for best
yield and capacity
Attributes Parameters Issues
Sterility assurance Membrane pore size Large size of pDNA
Particulate reduction Membrane chemistry
Shear sensitivity of
pDNA
Filtration capacity and
flux
Driving force
Viscosity of pDNA
solution (especially
at high
concentration,
>5mg/mL)
pDNA yield Formulation
Bacterial retention
for adjuvanted pDNA
solutions
Filtration endpoint
47. • Internal data and literature search show that pDNA size,
purity, and concentration have significant effect on sterile
filtration
• Effective size of pDNA can be altered by changing the ionic
concentration of background buffer solution
• Yield issues (<90%) are common with >10kbp plasmid and
can be significant (<80%) with >20kbp
Sterile Filtration
pDNA Sterile Filtration Optimization Suggestions
Data driven strategies and considerations for scalable purification of Plasmid DNA for use in vaccine manufacturing47
Optimization
Parameter
Yield Capacity Product integrity
Salt concentration X X
Supercoiled pDNA
content (purity) X X
Filtration endpoint X
Membrane type – PVDF
or PES X – PES X - PES
pDNA concentration X X
Feed flux or pressure X
doi:10.1002/bit.21575