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.
Chemically ligated gRNAs for CRISPR applications.Minghong Zhong
lgRNA (Chemically ligated gRNA) is designed to improve the functions and selectivity of gRNA, particularly its efficacy, less off-target effect and stability, and the cost-effectiveness in its industrial production.
lgRNAs are very useful for therapeutic and biotechnological applications of CRISPR, particularly for multiplexing and genome scale screening with its convenience for constructing evenly distributed gRNA libraries, by bypassing carcinogenesis and immunogenicity, limits in packaging capacities, random integration into host genomes, and the complicacy and tissue infection tropisms of viral vectors and plasmids.
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying the genomes of organisms ranging from E. coli to humans. In this presentation, we discuss various methods for generating the crRNA and tracrRNA components that are required for guiding the Cas9 endonuclease to genomic targets. You will also learn how to optimize a new 2-part CRISPR RNA system from IDT that offers multiple benefits over other technologies.
CRISPR has become an increasingly popular tool for genome editing, in part because it is highly flexible and relatively easy to implement compared to other technologies. However, for scientists beginning to work with this method, the wide range of products and variety of editing approaches can be overwhelming. In this presentation, Justin Barr provides a simple explanation of the steps for planning your experiment, including guide RNA design, an overview of delivery methods, and options for measuring editing results. He also discusses how to generate specific mutations in the genome using homology-directed repair (HDR).
Advancing Microbiome Research: From challenging samples to insight with Confi...QIAGEN
Microbiome research encompasses sample types as diverse as the human gut, Antarctic soil, ocean water and acidic hot spring biofilms. These samples are challenging because they are difficult to lyse, with some microbes containing a tough extracellular matrix. Incomplete lysis of a microbial community results in an inaccurate representation of the microbial content of the sample. Additionally, PCR inhibitors present in these samples, especially humic acids, polysaccharides, polyphenolics, lipids and heavy metals result in inaccurate quantification of nucleic acids that may inhibit downstream applications such as qPCR and NGS.
Chemically ligated gRNAs for CRISPR applications.Minghong Zhong
lgRNA (Chemically ligated gRNA) is designed to improve the functions and selectivity of gRNA, particularly its efficacy, less off-target effect and stability, and the cost-effectiveness in its industrial production.
lgRNAs are very useful for therapeutic and biotechnological applications of CRISPR, particularly for multiplexing and genome scale screening with its convenience for constructing evenly distributed gRNA libraries, by bypassing carcinogenesis and immunogenicity, limits in packaging capacities, random integration into host genomes, and the complicacy and tissue infection tropisms of viral vectors and plasmids.
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying the genomes of organisms ranging from E. coli to humans. In this presentation, we discuss various methods for generating the crRNA and tracrRNA components that are required for guiding the Cas9 endonuclease to genomic targets. You will also learn how to optimize a new 2-part CRISPR RNA system from IDT that offers multiple benefits over other technologies.
CRISPR has become an increasingly popular tool for genome editing, in part because it is highly flexible and relatively easy to implement compared to other technologies. However, for scientists beginning to work with this method, the wide range of products and variety of editing approaches can be overwhelming. In this presentation, Justin Barr provides a simple explanation of the steps for planning your experiment, including guide RNA design, an overview of delivery methods, and options for measuring editing results. He also discusses how to generate specific mutations in the genome using homology-directed repair (HDR).
Advancing Microbiome Research: From challenging samples to insight with Confi...QIAGEN
Microbiome research encompasses sample types as diverse as the human gut, Antarctic soil, ocean water and acidic hot spring biofilms. These samples are challenging because they are difficult to lyse, with some microbes containing a tough extracellular matrix. Incomplete lysis of a microbial community results in an inaccurate representation of the microbial content of the sample. Additionally, PCR inhibitors present in these samples, especially humic acids, polysaccharides, polyphenolics, lipids and heavy metals result in inaccurate quantification of nucleic acids that may inhibit downstream applications such as qPCR and NGS.
DNA Methylation: An Essential Element in Epigenetics Facts and TechnologiesQIAGEN
Check out this slide deck from Dr. Thorsten Singer and Dr. Ralf Peist to learn about DNA methylation in epigenetics, from its significance in cancer to strategies for studying it.
Developing a Scalable Upstream Bioreactor Process for Lentiviral Vector Produ...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3b3Jc77
Gene therapies hold the promise to change lives. As your path to patients accelerates, how can you assure the robust process design, intensification and scalability that meets your evolving manufacturing needs? What benefits can a templated process bring to your commercial success?
As gene therapy progresses toward broader clinical and commercial success, the industry is shifting from treating rare conditions to those of larger populations. This requires scalable solutions for process intensification. In this webinar, we’ll discuss scale-up development for a common viral vector in gene therapy, lentivirus, using the VirusExpress™ Lentiviral Production Platform in Mobius® single-use bioreactors. We will highlight critical considerations when moving from bench-scale to clinical scale process design with manufacturability in mind to ensure commercial readiness. Finally, we’ll review the significant benefits of implementing a templated manufacturing process.
In this webinar you will learn:
• Scale-up development of a suspension-based lentivirus production process
• Designing a process that is manufacturing-friendly and supports commercialization
• The benefits of having a templated manufacturing process
Application Note: A Simple One-Step Library Prep Method To Enable AmpliSeq Pa...QIAGEN
Targeted amplicon sequencing is a cost-effective, convenient and rapid method for variant detection. This application note outlines a straightforward workflow that uses the QIAseq 1-Step Amplicon Library kit to verify AmpliSeq targeted sequencing assays on the Illumina sequencing instruments. By combining end-repair and ligation, the QIAseq 1-Step Amplicon Library Kit offers a fast and efficient 30-minute procedure for the preparation of high-quality, artifact-free Illumina libraries from any PCR amplicons, including AmpliSeq Panels.
Recent breakthroughs in genome editing technology have led to a rapid adoption that parallels that seen with RNAi. And like RNAi, these methods are taking the scientific world by storm, with high profile publications in fields as diverse as HIV treatment, stem cell therapy, food crop modification and drug development to name but a few.
Critically, the endogenous modification of genes enables the study of their function in a physiological context. It also overcomes some of the artefacts that can result from established techniques such as transgenesis and RNAi, which have mislead researchers with false positives or negatives. Until recently however genome editing required considerable technical expertise, and consequently was a relatively niche pursuit.
In this talk we will look at how the latest developments in genome editing tools have changed this, with improvements in both ease-of-use and targeting efficiency, as well as a concomitant reduction in costs opening up these approaches to the wider scientific community.
Rapid adoption of the CRISPR/Cas9 system has for example led to a long list of organisms and tissues in which genetic changes have been made with high efficiency. Other technologies such as recombinant adeno-associated virus (rAAV) offer further precision, stimulating the cell’s high-fidelity DNA repair pathways to insert exogenous sequence with unrivalled specificity. Targeting efficiency can be improved still further by using the technologies in combination – genome cutting induced by CRISPR can significantly enhance homologous recombination mediated by rAAV.
Despite these rapid advances, some pitfalls remain, and so we’ll discuss some of the key considerations for avoiding these, ranging from simply picking the right tool for the job to designing an experiment that maximises chances of success.
Finally we’ll look at how genome editing is being applied to both basic and translational research, and in both a gene-specific and genome wide manner. For the study of disease associated genes and mutations scientists can now complement wide panels of tumour cells with genetically defined isogenic cell pairs identical in all but precise modifications in their gene of interest. The ease-of-design and efficiency of the CRISPR system is also being exploited for genome wide synthetic lethality screens, facilitating rapid drug target identification with significantly reduced risk of false negatives and off-target false positives. And again, further synergies are achieved when these approaches are combined to look for potential synthetic lethal targets in specific genomic contexts.
GENESIS™: Comprehensive genome editing - Translating genetic information into personalised medicines.
Horizon is the only source of rAAV expertise and is uniquely capable of exploiting multiple platforms: CRISPR, ZFNs and rAAV singularly or combined. Horizon’s scientists are experts at all forms of gene editing and so have the experience to help guide customers towards the approach that best suits their project
How to reach High Plasma Protein Concentration with Single-Pass TFFMerck Life Sciences
This webinar will discuss our collaboration with Takeda on the development of a single-pass TFF system as an alternative to traditional TFF for concentrating a plasma-derived IgG solution.
Single-Pass Tangential Flow Filtration (SPTFF) is a technology that requires only one pass through the filter assembly to achieve the desired concentration with no recirculation of product.
SPTFF can offer many advantages in downstream processing, such as:
• Increased capacity and reduced process time
• Increased yield and product recovery
• Optimized processing of highly shear-sensitive products
• Reduced foam formation
• Reduced cost of goods
This presentation will cover our collaboration with Takeda, formerly Shire, for the development of a specific SPTFF system as an alternative to traditional TFF for concentrating a plasma-derived Immunoglobulin G (IgG) solution from 10% to 20%. Due to promising results, plans are underway to replace the currently used batch TFF process with a SPTFF step.
In this webinar, we will discuss:
- A comparison of traditional TFF versus SPTFF
- Design of Experiments (DOE) approach toward initial process development work and determination of the optimal parameters
- Process run results, including final product yield and product quality
The availability of affordable, high quality, custom gene synthesis has greatly expanded what is possible for labs in numerous research areas. IDT offers a variety of gene synthesis solutions, including our revolutionary double-stranded gBlocks Gene Fragments. In this presentation, Dr Adam Clore discusses the many applications of gBlocks Gene Fragments, such as controls for qPCR and next generation sequencing, and donor templates for homology directed repair in CRISPR experiments. Learn more at www.idtdna.com/gblocks
My talk for the International Genomics session at ABRF 2017. Describing the issues caused by the uncontrolled naming of NGS methods: some examples and some suggestions about how to fix this.
New technology and workflow for integrated collection, stabilization and puri...QIAGEN
Research into non-invasive prenatal testing (NIPT) and circulating tumor DNA (ctDNA) testing based on circulating cell-free DNA (ccfDNA) is rapidly expanding. However, detection and quantification of ccfDNA is compromised by the release of genomic DNA (gDNA) from lymphocytes due to mechanical lysis or apoptosis during blood collection, storage and transport. PreAnalytiX has developed the PAXgene Blood ccfDNA System, consisting of the PAXgene Blood ccfDNA Tube, a plastic blood collection tube with a unique, non-crosslinking chemistry that preserves extracellular levels of ccfDNA and prevents the release of intracellular DNA from cells into the plasma, and the QIAsymphonyPAXgene Blood ccfDNA Kit for automated ccfDNA extraction from up to 5 ml of plasma. In this slidedeck, this new technology development is presented in comparison to other existing technologies.
Dr Adam Clore discusses uses for gBlocks® Gene Fragments in the context of the 2015 iGEM competition. Dr Clore also describes how iGEM teams can register to receive 20 kb of free gBlocks Gene Fragments for their projects.
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.
Overcome the challenges of Nucleic acid isolation from PCR inhibitor-rich mic...QIAGEN
This presentation will focus on nucleic acid extraction tools developed by QIAGEN that facilitate accurate non-biased community analysis and eliminate common amplification problems via the depletion of endogenous polymerase inhibitors using our patented Inhibitor Removal Technology.
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying genomes of organisms ranging from E. coli to humans. Additionally, the simple gene targeting mechanism of CRISPR technology has been modified and adapted to other applications that include gene regulation, detection of intercellular trafficking, and pathogen detection. With a wealth of methods for introducing Cas9 and gRNAs into cells, it can be challenging to decide where to start. In this presentation, Dr Adam Clore describes the CRISPR mechanism and some of the most prominent uses for CRISPR, along with methods where IDT technologies can assist scientists in designing, testing, and executing a variety of CRISPR-mediated experiments. For more informaton, visit: http://www.idtdna.com/crispr
Biosafety in Gene Therapy: Applying the latest regulatory guidance for RCL te...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/33WUiqE
Ensuring the safety and quality of your lentiviral vector is of the utmost importance. Attend this webinar to learn about testing strategies to monitor replication competent lentivirus. You will also hear about recent changes in regulatory guidance with regards to sample types and volumes tested.
The use of lentivirus vectors to produce groundbreaking gene therapies is on the rise. Ensuring the biosafety and quality of these vectors is achieved through a multi-tiered testing approach.
For lentivirus-based therapies, generation of replication competent particles is a potential risk. While improvements in design and manufacturing have decreased the probability of producing replication competent viruses, regulatory agencies provide guidelines to test for their presence at multiple stages in production. This webinar reviews the strategies for monitoring replication competent lentiviruses. We describe current methods and address: Sample types, testing volumes, and expected results.
In this webinar, you will learn about:
• The latest FDA regulatory guidelines on replication competent lentivirus (RCL) testing
• Methods used to monitor RCL
• Considerations on sample type and volume requirements
The Role of BPOG Extractables Data in the Effective Adoption of Single-Use Sy...Merck Life Sciences
The successful adoption of single-use technologies in a biopharmaceutical process largely relies on confidently selecting the right components for use in the fluid path of a product, within a specific process. An important step in choosing such components requires generating an extractables profile, which can be done by carefully selecting the solvent streams and extraction conditions to model the product and process steps complemented with the right analytical strategy.
In this webinar, you will learn:
● An approach to adopt the BioPhorum Operations Group (BPOG) extractables protocol as a baseline testing strategy.
● How to apply extractables data to a specific process followed by a systematic, risk-based safety assessment approach used for comparing known safety concern thresholds.
● The important stages in the risk assessment process as demonstrated by case studies from typical drug manufacturing processes where single-use components were used.
Serum-free Media for Therapeutic Cell Manufacturing – Challenges and InnovationsMerck Life Sciences
The need for high quality materials that are animal origin-free and compatible with a limited number of downstream processing steps will increase as cell therapies achieve clinical success. Large scale manufacturing necessitates transition from planar culture to technologies such as stirred tank bioreactors where culture of suspension cells or adherent-dependent cells on microcarriers is enabled.
This webinar will discuss challenges and solutions to the elimination of animal-derived components from cell culture processes, with focus on human mesenchymal stromal/stem cells (hMSCs). Fetal bovine serum in particular is associated with regulatory, supply, and consistency challenges, yet a wide range of performance has been observed between different serum-free media formulations for expansion of hMSCs in planar formats. Moreover, a positive performance in static culture is not necessarily predictive of that under agitated conditions with microcarriers, highlighting ongoing challenges to the generation of a fully chemically-defined and scalable cell culture medium. Through use of pharma-grade basal media manufactured with advanced milling technology and EMPROVE® raw materials, as well as transition to serum-free supplementation and process development activities, the robust expansion of hMSCs across platforms has been achieved.
Presented by Aletta Schnitzler, Senior Scientist on 5/5/16
DNA Methylation: An Essential Element in Epigenetics Facts and TechnologiesQIAGEN
Check out this slide deck from Dr. Thorsten Singer and Dr. Ralf Peist to learn about DNA methylation in epigenetics, from its significance in cancer to strategies for studying it.
Developing a Scalable Upstream Bioreactor Process for Lentiviral Vector Produ...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3b3Jc77
Gene therapies hold the promise to change lives. As your path to patients accelerates, how can you assure the robust process design, intensification and scalability that meets your evolving manufacturing needs? What benefits can a templated process bring to your commercial success?
As gene therapy progresses toward broader clinical and commercial success, the industry is shifting from treating rare conditions to those of larger populations. This requires scalable solutions for process intensification. In this webinar, we’ll discuss scale-up development for a common viral vector in gene therapy, lentivirus, using the VirusExpress™ Lentiviral Production Platform in Mobius® single-use bioreactors. We will highlight critical considerations when moving from bench-scale to clinical scale process design with manufacturability in mind to ensure commercial readiness. Finally, we’ll review the significant benefits of implementing a templated manufacturing process.
In this webinar you will learn:
• Scale-up development of a suspension-based lentivirus production process
• Designing a process that is manufacturing-friendly and supports commercialization
• The benefits of having a templated manufacturing process
Application Note: A Simple One-Step Library Prep Method To Enable AmpliSeq Pa...QIAGEN
Targeted amplicon sequencing is a cost-effective, convenient and rapid method for variant detection. This application note outlines a straightforward workflow that uses the QIAseq 1-Step Amplicon Library kit to verify AmpliSeq targeted sequencing assays on the Illumina sequencing instruments. By combining end-repair and ligation, the QIAseq 1-Step Amplicon Library Kit offers a fast and efficient 30-minute procedure for the preparation of high-quality, artifact-free Illumina libraries from any PCR amplicons, including AmpliSeq Panels.
Recent breakthroughs in genome editing technology have led to a rapid adoption that parallels that seen with RNAi. And like RNAi, these methods are taking the scientific world by storm, with high profile publications in fields as diverse as HIV treatment, stem cell therapy, food crop modification and drug development to name but a few.
Critically, the endogenous modification of genes enables the study of their function in a physiological context. It also overcomes some of the artefacts that can result from established techniques such as transgenesis and RNAi, which have mislead researchers with false positives or negatives. Until recently however genome editing required considerable technical expertise, and consequently was a relatively niche pursuit.
In this talk we will look at how the latest developments in genome editing tools have changed this, with improvements in both ease-of-use and targeting efficiency, as well as a concomitant reduction in costs opening up these approaches to the wider scientific community.
Rapid adoption of the CRISPR/Cas9 system has for example led to a long list of organisms and tissues in which genetic changes have been made with high efficiency. Other technologies such as recombinant adeno-associated virus (rAAV) offer further precision, stimulating the cell’s high-fidelity DNA repair pathways to insert exogenous sequence with unrivalled specificity. Targeting efficiency can be improved still further by using the technologies in combination – genome cutting induced by CRISPR can significantly enhance homologous recombination mediated by rAAV.
Despite these rapid advances, some pitfalls remain, and so we’ll discuss some of the key considerations for avoiding these, ranging from simply picking the right tool for the job to designing an experiment that maximises chances of success.
Finally we’ll look at how genome editing is being applied to both basic and translational research, and in both a gene-specific and genome wide manner. For the study of disease associated genes and mutations scientists can now complement wide panels of tumour cells with genetically defined isogenic cell pairs identical in all but precise modifications in their gene of interest. The ease-of-design and efficiency of the CRISPR system is also being exploited for genome wide synthetic lethality screens, facilitating rapid drug target identification with significantly reduced risk of false negatives and off-target false positives. And again, further synergies are achieved when these approaches are combined to look for potential synthetic lethal targets in specific genomic contexts.
GENESIS™: Comprehensive genome editing - Translating genetic information into personalised medicines.
Horizon is the only source of rAAV expertise and is uniquely capable of exploiting multiple platforms: CRISPR, ZFNs and rAAV singularly or combined. Horizon’s scientists are experts at all forms of gene editing and so have the experience to help guide customers towards the approach that best suits their project
How to reach High Plasma Protein Concentration with Single-Pass TFFMerck Life Sciences
This webinar will discuss our collaboration with Takeda on the development of a single-pass TFF system as an alternative to traditional TFF for concentrating a plasma-derived IgG solution.
Single-Pass Tangential Flow Filtration (SPTFF) is a technology that requires only one pass through the filter assembly to achieve the desired concentration with no recirculation of product.
SPTFF can offer many advantages in downstream processing, such as:
• Increased capacity and reduced process time
• Increased yield and product recovery
• Optimized processing of highly shear-sensitive products
• Reduced foam formation
• Reduced cost of goods
This presentation will cover our collaboration with Takeda, formerly Shire, for the development of a specific SPTFF system as an alternative to traditional TFF for concentrating a plasma-derived Immunoglobulin G (IgG) solution from 10% to 20%. Due to promising results, plans are underway to replace the currently used batch TFF process with a SPTFF step.
In this webinar, we will discuss:
- A comparison of traditional TFF versus SPTFF
- Design of Experiments (DOE) approach toward initial process development work and determination of the optimal parameters
- Process run results, including final product yield and product quality
The availability of affordable, high quality, custom gene synthesis has greatly expanded what is possible for labs in numerous research areas. IDT offers a variety of gene synthesis solutions, including our revolutionary double-stranded gBlocks Gene Fragments. In this presentation, Dr Adam Clore discusses the many applications of gBlocks Gene Fragments, such as controls for qPCR and next generation sequencing, and donor templates for homology directed repair in CRISPR experiments. Learn more at www.idtdna.com/gblocks
My talk for the International Genomics session at ABRF 2017. Describing the issues caused by the uncontrolled naming of NGS methods: some examples and some suggestions about how to fix this.
New technology and workflow for integrated collection, stabilization and puri...QIAGEN
Research into non-invasive prenatal testing (NIPT) and circulating tumor DNA (ctDNA) testing based on circulating cell-free DNA (ccfDNA) is rapidly expanding. However, detection and quantification of ccfDNA is compromised by the release of genomic DNA (gDNA) from lymphocytes due to mechanical lysis or apoptosis during blood collection, storage and transport. PreAnalytiX has developed the PAXgene Blood ccfDNA System, consisting of the PAXgene Blood ccfDNA Tube, a plastic blood collection tube with a unique, non-crosslinking chemistry that preserves extracellular levels of ccfDNA and prevents the release of intracellular DNA from cells into the plasma, and the QIAsymphonyPAXgene Blood ccfDNA Kit for automated ccfDNA extraction from up to 5 ml of plasma. In this slidedeck, this new technology development is presented in comparison to other existing technologies.
Dr Adam Clore discusses uses for gBlocks® Gene Fragments in the context of the 2015 iGEM competition. Dr Clore also describes how iGEM teams can register to receive 20 kb of free gBlocks Gene Fragments for their projects.
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.
Overcome the challenges of Nucleic acid isolation from PCR inhibitor-rich mic...QIAGEN
This presentation will focus on nucleic acid extraction tools developed by QIAGEN that facilitate accurate non-biased community analysis and eliminate common amplification problems via the depletion of endogenous polymerase inhibitors using our patented Inhibitor Removal Technology.
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying genomes of organisms ranging from E. coli to humans. Additionally, the simple gene targeting mechanism of CRISPR technology has been modified and adapted to other applications that include gene regulation, detection of intercellular trafficking, and pathogen detection. With a wealth of methods for introducing Cas9 and gRNAs into cells, it can be challenging to decide where to start. In this presentation, Dr Adam Clore describes the CRISPR mechanism and some of the most prominent uses for CRISPR, along with methods where IDT technologies can assist scientists in designing, testing, and executing a variety of CRISPR-mediated experiments. For more informaton, visit: http://www.idtdna.com/crispr
Biosafety in Gene Therapy: Applying the latest regulatory guidance for RCL te...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/33WUiqE
Ensuring the safety and quality of your lentiviral vector is of the utmost importance. Attend this webinar to learn about testing strategies to monitor replication competent lentivirus. You will also hear about recent changes in regulatory guidance with regards to sample types and volumes tested.
The use of lentivirus vectors to produce groundbreaking gene therapies is on the rise. Ensuring the biosafety and quality of these vectors is achieved through a multi-tiered testing approach.
For lentivirus-based therapies, generation of replication competent particles is a potential risk. While improvements in design and manufacturing have decreased the probability of producing replication competent viruses, regulatory agencies provide guidelines to test for their presence at multiple stages in production. This webinar reviews the strategies for monitoring replication competent lentiviruses. We describe current methods and address: Sample types, testing volumes, and expected results.
In this webinar, you will learn about:
• The latest FDA regulatory guidelines on replication competent lentivirus (RCL) testing
• Methods used to monitor RCL
• Considerations on sample type and volume requirements
The Role of BPOG Extractables Data in the Effective Adoption of Single-Use Sy...Merck Life Sciences
The successful adoption of single-use technologies in a biopharmaceutical process largely relies on confidently selecting the right components for use in the fluid path of a product, within a specific process. An important step in choosing such components requires generating an extractables profile, which can be done by carefully selecting the solvent streams and extraction conditions to model the product and process steps complemented with the right analytical strategy.
In this webinar, you will learn:
● An approach to adopt the BioPhorum Operations Group (BPOG) extractables protocol as a baseline testing strategy.
● How to apply extractables data to a specific process followed by a systematic, risk-based safety assessment approach used for comparing known safety concern thresholds.
● The important stages in the risk assessment process as demonstrated by case studies from typical drug manufacturing processes where single-use components were used.
Serum-free Media for Therapeutic Cell Manufacturing – Challenges and InnovationsMerck Life Sciences
The need for high quality materials that are animal origin-free and compatible with a limited number of downstream processing steps will increase as cell therapies achieve clinical success. Large scale manufacturing necessitates transition from planar culture to technologies such as stirred tank bioreactors where culture of suspension cells or adherent-dependent cells on microcarriers is enabled.
This webinar will discuss challenges and solutions to the elimination of animal-derived components from cell culture processes, with focus on human mesenchymal stromal/stem cells (hMSCs). Fetal bovine serum in particular is associated with regulatory, supply, and consistency challenges, yet a wide range of performance has been observed between different serum-free media formulations for expansion of hMSCs in planar formats. Moreover, a positive performance in static culture is not necessarily predictive of that under agitated conditions with microcarriers, highlighting ongoing challenges to the generation of a fully chemically-defined and scalable cell culture medium. Through use of pharma-grade basal media manufactured with advanced milling technology and EMPROVE® raw materials, as well as transition to serum-free supplementation and process development activities, the robust expansion of hMSCs across platforms has been achieved.
Presented by Aletta Schnitzler, Senior Scientist on 5/5/16
Considerations for Manufacturing Commercial Antibody Drug Conjugates (ADCs) -...Merck Life Sciences
ADC manufacturing presents a unique set of challenges as compared to the well established practices employed for more traditional biologic products.
The development and commercialization of key intermediates, complex small-molecule APIs and biologic drug substances shouldn’t be such a headache. The uniqueness, versatility, and complexity involved in each project only means you need to make sure you’re well informed when it comes to the dos and don'ts of manufacturing commercial Antibody Drug Conjugates (ADCs). Tune in to hear a CMO perspective for an in-depth understanding on the subject of manufacturing commercial ADCs. Our team will discuss all of the considerations that must be addressed to successfully manufacture ADCs.
In this webinar you will learn:
- Facility Design and Cleaning Validation
- Advantages of Single Use Systems
- Process Control and Regulatory Strategies
Discover solutions for all phases of product development for genetox assessment from in silico analysis, screening, mode of action assessment, or GLP regulatory required assays. Our BioReliance® Genetic Toxicology Services director will share specifics and rationale for each assay category.
In this webinar you will:
- Learn the required regulatory assays
- Understand why each assay is used and how to employ different assay designs
- Learn different assays and techniques to screen potential compounds and understand mechanism and mode of action
Presented by Rohan Kulkarni, Ph.D., ERT, Director Toxicology, Study Management on February 9, 2017
Modern BioManufacturing: Single-Use Technologies in Configurable, Prefabricat...Merck Life Sciences
A co-webinar describing a solution to biopharma's challenge of rapidly and rationally expanding capacity by employing single-use technologies, a templated process train, and pre-fabricated mobile/modular cleanrooms.
Biopharmaceutical companies on the verge of investing into manufacturing or facilities expansion face many questions and challenges. Speed, agility, and flexibility are becoming more critical to executing their changing production and distribution strategies. Platform facility designs which integrate the latest process technologies within innovative pre-fabricated cleanrooms are critical for addressing the trending desire to implement 'clonable' modular facilities that can be delivered in a timely fashion across multiple locations. Companies like Merck KGaA, Darmstadt, Germany and G-CON Manufacturing are working together to combine their technologies and develop simple yet robust platform solutions for industry.
As bioprocessing technologies intensify performance, volumetric requirements become less. As such, 2000L single-use bioreactors - or multiple bioreactors of similar or less volumes - now suffice for the production of novel or biosimilar recombinant proteins. Such a shift in the industry enables the development of more mobile, modular facility designs. We will describe the rationale for this collaboration and its result: a turn-key solution that integrates a templated process train with a rapidly-deployable facility platform. By combining the unique advantages found with the G-CON POD construction and the bioprocess technology expertise from within Merck KGaA, Darmstadt, Germany, the goal of creating a cost-effective, pre-fabricated alternative to historical 'stick built' facilities is being achieved. Additionally, the flexibility inherent to our approach provides for a greater configurability that confers more user-specified choice into the selection of options. Simple in concept, this solution is also robust, cost-effective, and conducive to tight timelines for implementation.
In this webinar you will learn:
- Basic options for facilities/capacity expansion
- The value of templated process trains employing single-use equipment
- How modular, prefabricated PODs® outfitted with such single-use bioprocessing equipment represent an attractive, cost-effective strategy for capacity expansion
POD® is a registered trademark of G-CON Manufacturing, Inc.
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
Are you involved with planning tech transfer of your drug product? Join this webinar to learn more about the regulations and considerations you need to consider and learnings from a case study.
According to ICH Q10, “The goal of technology transfer activities is to transfer product and process knowledge between development and manufacturing, and within or between manufacturing sites to achieve product realization. This knowledge forms the basis for the manufacturing process, control strategy, process validation approach, and ongoing continual improvement.”
As a result, there is an expectation for transfers to be performed in an organized, methodical manner with appropriate documentation. It is also expected that they happen between one Process Development group to another or to a Pilot Lab, from Process Development lab to clinical or commercial manufacturing, or from Process Development to external clinical manufacturing. Lastly, they may also happen between two company facilities at commercial scale, or between a company and an external contract manufacturing at commercial scale.
This presentation will cover points to consider for successful tech transfers with a focus on cGMP training requirements, and include lesson learned from real cases.
Presented by Guillaume Plane on September 22, 2016
Integrity testing is a critical operation, especially for sterilizing grade filters used in biopharmaceutical processing. When performed correctly, an integrity test is a fast, definitive, non-destructive way to assure filter retention performance. Fortunately, there are few ways a non-integral filter will pass the integrity test, eliminating the possibility a non-retentive filter is used undetected. Unfortunately, there are a lot of ways an integral filter can fail the integrity test, resulting in retests, lost time, productivity and potentially lost product.
In this webinar you will:
- Gain confidence in your integrity testing results
- Provide justification for retests
- Understand specific challenges and eliminate them to assure the integrity test can be performed correctly the first time
Rapid Methodologies for Biosafety Testing of Biologic TherapeuticsMerck Life Sciences
Learn about existing and emerging methods to accelerate biosafety testing of biologic therapies.
Speed to market for biologic therapeutics is ever more critical. However, the critical safety tests for these molecules, for example screening for adventitious agents such as viral contaminants, can be time consuming as well as challenging and laborious. Join us for this webinar as we explore how rapid methodologies are being used to not only accelerate this process, but also enhance quality by reducing testing complexity. Existing technologies as well as emerging trends will be discussed, along with the implications these may have on the regulatory landscape.
In this webinar you will learn:
● Which existing and emerging technologies are having now, and will have in the future, an impact on biosaftey testing.
● The benefits as well as risks of employing rapid methods for biosafety screening.
● How the regulatory agencies are reacting to rapid testing methods as alternatives to existing methods.
Our Life Science business is fully dedicated to supporting small, biotech companies with cutting edge technologies. Besides technical aspects of molecule development and production, fundraising is omnipresent. This webinar will provide insights and perspectives from Merck Ventures, BV, a subsidiary of
Merck KGaA, Darmstadt, Germany.
At Merck Ventures, BV, a subsidiary of Merck KGaA, Darmstadt, Germany, the strategic corporate venture capital arm of Merck KGaA, Darmstadt Germany, we drive innovation and back entrepreneurs through equity investments and hands-on support. We focus on areas that impact the vitality and sustainability of our current and future businesses.
This webinar will provide you with the ABCs of venture capital including:
• How venture capital works
• The role of a corporate venture capital
• How we look at opportunities
Time is of the Essence: Creating a New Synergy Between Single-Use Adopters an...Merck Life Sciences
Biopharmaceutical companies race to achieve milestones, advance promising molecules, improve productivity and reduce costs. In addition to perfecting the biology, companies must grapple with challenges such as standardization of process technology, supply security and process economics, especially in light of the burgeoning interest and adoption of single-use systems. Efficient implementation of single-use systems is more than incorporating disposable components into the process. It requires a new approach by suppliers of these systems to nimbly and effectively address such challenges.
This session will highlight best practices for creating a new synergy between biopharmaceutical companies and single-use suppliers to strike the right balance of design flexibility, supply predictability and reliable lead times necessary to beat the clock. A case study describing a more efficient and practical “bottoms-up” approach to configuring and delivering single-use assemblies will be presented.
In this webinar, you will learn:
- How to choose single-use assemblies that are best suited for your process
- Risk reduction strategies that incorporate stock or specialized single-use assemblies vs. custom solutions
- Best practices to strike the right balance of design flexibility and supply predictability
Expression and purification of recombinant proteins in Bacterial and yeast sy...Shreya Feliz
This presentation gives the information about bacterial and yeast system as host for expressing recombinant proteins, suitable vectors, strains of host, Pros and cons of this system, different purification techniques and commercially available proteins produced so far by this system.
Recombinant baculoviruses are widely used to
express heterologous genes in cultured insect cells
and insect larvae. For large-scale applications, the
baculovirus expression vector system (BEVS) is particularly
advantageous.
Viral Risk Mitigation Strategies: Key Considerations in the Prevention and De...Merck Life Sciences
Regulatory guidelines have defined industry best practices around adventitious virus contamination and risk mitigation in terms of patient safety.
Today, the industry is taking a closer look at minimizing the business risk associated with viral contamination and is taking a more directed view of risk mitigation. This approach includes virus prevention and detection, in addition to removal.
From cell culture seed train to final fill vial, this presentation will describe:
-Potential risks associated with different areas of biotech processes
-What can be done to minimize adventitious virus risk in those areas.
The overarching strategy of risk mitigation will include evaluation of raw materials, modified expression systems, environmental controls, upstream and downstream processing, as well as testing and regulatory considerations.
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
Accelerate Delivery of High Producing Cell LinesMilliporeSigma
Watch the interactive recording here: https://bit.ly/30FTDG0
The quest for a viable upstream process relies on generation of a cell line expressing the protein of interest. Unfortunately, the search for the best-producing clone is often compared with looking for a needle in a haystack. Making this more challenging is the pressure to get it right the first time, quickly and while mitigating risk and costs.
Although a lot of efforts are made on the clonal selection, there is often few to none optimization done on the expression cassette, including promoter and enhancer selection, or signal peptide. The statistical approach on how many clones should be screened to get to a good producer is often overlooked as well.
We combined a new generation of promoters and enhancers to improve strategies on pool and mini pool screening with both CHO-K1 and our own CHOZN® GS which helped deliver high-producing clones in an accelerated timeline. In addition, we are able to begin process development in parallel with cell line development, further reducing timelines.
In this webinar, you will learn:
* How the strategy approach can help reducing the overall timeline of cell line generation
* How we have expanded our platform by designing a completely new vector/cell/process template
* How we have worked on promoters, enhancers, pool/mini-pool approach as well as on timelines from DNA to clone
Watch the interactive recording here: https://bit.ly/30FTDG0
The quest for a viable upstream process relies on generation of a cell line expressing the protein of interest. Unfortunately, the search for the best-producing clone is often compared with looking for a needle in a haystack. Making this more challenging is the pressure to get it right the first time, quickly and while mitigating risk and costs.
Although a lot of efforts are made on the clonal selection, there is often few to none optimization done on the expression cassette, including promoter and enhancer selection, or signal peptide. The statistical approach on how many clones should be screened to get to a good producer is often overlooked as well.
We combined a new generation of promoters and enhancers to improve strategies on pool and mini pool screening with both CHO-K1 and our own CHOZN® GS which helped deliver high-producing clones in an accelerated timeline. In addition, we are able to begin process development in parallel with cell line development, further reducing timelines.
In this webinar, you will learn:
* How the strategy approach can help reducing the overall timeline of cell line generation
* How we have expanded our platform by designing a completely new vector/cell/process template
* How we have worked on promoters, enhancers, pool/mini-pool approach as well as on timelines from DNA to clone
HIV Vaccines Process Development & Manufacturing - Pitfalls & PossibilitiesKBI Biopharma
Originally presented at the HIV Vaccine Manufacturing Workshop –July 19th& 20th, 2017 by Abhinav A. Shukla, Ph.D.Senior Vice PresidentDevelopment & ManufacturingKBI Biopharma, Durham NC
Learn more about the Valitacell fluorescent polarisation based IgG quantification assay 'ValitaTITER' and about our novel ChemStress fingerprinting assay for cell line development. For more information about our products and pricing, please contact info@valitacell.com
Cell Line Development: Reducing timelines and increasing titres fujifilmdiosynth
Cell line development: Reducing timelines and increasing titres by identification of host cell lines with improved characteristics. To develop a mammalian expression platform which rapidly leads to efficient, robust and high quality biomanufacturing processes
Streamlining Biopharmaceutical Cell Line Development - Reducing risk and decr...MilliporeSigma
CHO cells with their unique characteristics, represent the major expression system within the biopharmaceutical industry. However, one of the major challenges in cell line development is to identify those rare, high-producing clones in a huge population of non-expressing or low-expressing cell lines. This leads to laborious and time consuming cell line development processes. This webinar will educate the audience about challenges faced with traditional expression systems and how the CHO cell line with the glutamine synthethase knock-out via Zinc Finger Nucleases provides benefits for fast and efficient cell line development as well as stable and high titer expression. We will explore additional cell line engineering targets that can be modified to engineer a cell line that mitigates risks and removes bottlenecks throughout the biopharmaceutical process.
In this webinar, you will learn:
• What are the benefits of using an optimized/engineered expression system?
• What can be done throughout the cell line development process to mitigate risks and remove bottlenecks?
• Applications of cell line engineering for further upstream biopharmaceutical enhancements.
Streamlining Biopharmaceutical Cell Line Development - Reducing risk and decr...Merck Life Sciences
CHO cells with their unique characteristics, represent the major expression system within the biopharmaceutical industry. However, one of the major challenges in cell line development is to identify those rare, high-producing clones in a huge population of non-expressing or low-expressing cell lines. This leads to laborious and time consuming cell line development processes. This webinar will educate the audience about challenges faced with traditional expression systems and how the CHO cell line with the glutamine synthethase knock-out via Zinc Finger Nucleases provides benefits for fast and efficient cell line development as well as stable and high titer expression. We will explore additional cell line engineering targets that can be modified to engineer a cell line that mitigates risks and removes bottlenecks throughout the biopharmaceutical process.
In this webinar, you will learn:
• What are the benefits of using an optimized/engineered expression system?
• What can be done throughout the cell line development process to mitigate risks and remove bottlenecks?
• Applications of cell line engineering for further upstream biopharmaceutical enhancements.
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
Data driven strategies and considerations for scalable purification of Plasmi...Merck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/2JeT1U9
Plasmid DNA (pDNA) presents unique manufacturing challenges. While research scale purification kits simplify small pDNA preparations, scalable manufacturing must leverage significant process understanding. This webinar presents scalable solutions for all downstream unit operations from harvest to bulk filtration.
Plasmid DNA (pDNA) has been an important scientific tool for decades, but as clinical and commercial applications increase, manufacturers of pDNA face pressure to optimize production activities to meet demand while maintaining critical quality attributes. Key challenges in pDNA manufacturing exist around purification unit operations due to its large size, high viscosity, shear sensitivity, and similarities between pDNA and impurities. Overcoming downstream challenges with scalable techniques requires in depth knowledge of unit operation parameters and holistic process understanding. Our work investigates parameters and key considerations for purification unit operations including harvest, lysis, clarification, tangential flow filtration, chromatography, and sterilizing grade filtration.
In this webinar, you will learn:
• Parameters for E. coli harvest using microfiltration tangential flow filtration
• Key considerations for scalable alkaline lysis
• Filter selection guidance for clarification of alkaline lysate
• Purification strategies using AEX chromatography resins and membranes
• Implementation considerations for ultrafiltration/diafiltration
• Watch-outs for sterile filtration
• Purification process flow for Plasmid DNA
Cancer Research & the Challenges of FFPE Samples – An IntroductionQIAGEN
A cascade of complex genetic and epigenetic changes regulate tumor formation and progression. Gene expression analyses can shed light on these changes at a molecular level and identify the key genes and associated pathways involved in cancer. Often the samples used in cancer research are FFPE samples, which pose a significant challenge in terms of nucleic acid quality. The quality of nucleic acids extracted from FFPE samples depends on a number of factors, including how the samples were handled before, during and after fixation and embedding.
Dr. Vishwadeepak Tripathi describes the variability of sample purification from FFPE samples – in particular, samples to be used in cancer research. What are the challenges and solutions, and what quality control approach can ensure credible results? This webinar will focus on sample purification and the quality control of FFPE samples and compare different automated purification procedures.
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
Cellca is a leading provider of Cell Line Development Services allowing customers easy open access to a cost effective reliable technology platform consistently delivering well characterised stable research clones from DNA to Research Cell Bank (RCB) in 4 months with titres upwards of 3.0 g/L in an easily scalable fed batch process.
Similar to Delivering More Efficient Therapeutic Protein Expression Systems Through Cell Line Engineering Applications (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
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
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.
CHAPTER 1 SEMESTER V - ROLE OF PEADIATRIC NURSE.pdfSachin Sharma
Pediatric nurses play a vital role in the health and well-being of children. Their responsibilities are wide-ranging, and their objectives can be categorized into several key areas:
1. Direct Patient Care:
Objective: Provide comprehensive and compassionate care to infants, children, and adolescents in various healthcare settings (hospitals, clinics, etc.).
This includes tasks like:
Monitoring vital signs and physical condition.
Administering medications and treatments.
Performing procedures as directed by doctors.
Assisting with daily living activities (bathing, feeding).
Providing emotional support and pain management.
2. Health Promotion and Education:
Objective: Promote healthy behaviors and educate children, families, and communities about preventive healthcare.
This includes tasks like:
Administering vaccinations.
Providing education on nutrition, hygiene, and development.
Offering breastfeeding and childbirth support.
Counseling families on safety and injury prevention.
3. Collaboration and Advocacy:
Objective: Collaborate effectively with doctors, social workers, therapists, and other healthcare professionals to ensure coordinated care for children.
Objective: Advocate for the rights and best interests of their patients, especially when children cannot speak for themselves.
This includes tasks like:
Communicating effectively with healthcare teams.
Identifying and addressing potential risks to child welfare.
Educating families about their child's condition and treatment options.
4. Professional Development and Research:
Objective: Stay up-to-date on the latest advancements in pediatric healthcare through continuing education and research.
Objective: Contribute to improving the quality of care for children by participating in research initiatives.
This includes tasks like:
Attending workshops and conferences on pediatric nursing.
Participating in clinical trials related to child health.
Implementing evidence-based practices into their daily routines.
By fulfilling these objectives, pediatric nurses play a crucial role in ensuring the optimal health and well-being of children throughout all stages of their development.
Leading the Way in Nephrology: Dr. David Greene's Work with Stem Cells for Ki...Dr. David Greene Arizona
As we watch Dr. Greene's continued efforts and research in Arizona, it's clear that stem cell therapy holds a promising key to unlocking new doors in the treatment of kidney disease. With each study and trial, we step closer to a world where kidney disease is no longer a life sentence but a treatable condition, thanks to pioneers like Dr. David Greene.
QA Paediatric dentistry department, Hospital Melaka 2020Azreen Aj
QA study - To improve the 6th monthly recall rate post-comprehensive dental treatment under general anaesthesia in paediatric dentistry department, Hospital Melaka
How many patients does case series should have In comparison to case reports.pdfpubrica101
Pubrica’s team of researchers and writers create scientific and medical research articles, which may be important resources for authors and practitioners. Pubrica medical writers assist you in creating and revising the introduction by alerting the reader to gaps in the chosen study subject. Our professionals understand the order in which the hypothesis topic is followed by the broad subject, the issue, and the backdrop.
https://pubrica.com/academy/case-study-or-series/how-many-patients-does-case-series-should-have-in-comparison-to-case-reports/
The dimensions of healthcare quality refer to various attributes or aspects that define the standard of healthcare services. These dimensions are used to evaluate, measure, and improve the quality of care provided to patients. A comprehensive understanding of these dimensions ensures that healthcare systems can address various aspects of patient care effectively and holistically. Dimensions of Healthcare Quality and Performance of care include the following; Appropriateness, Availability, Competence, Continuity, Effectiveness, Efficiency, Efficacy, Prevention, Respect and Care, Safety as well as Timeliness.
One of the most developed cities of India, the city of Chennai is the capital of Tamilnadu and many people from different parts of India come here to earn their bread and butter. Being a metropolitan, the city is filled with towering building and beaches but the sad part as with almost every Indian city
Antibiotic Stewardship by Anushri Srivastava.pptxAnushriSrivastav
Stewardship is the act of taking good care of something.
Antimicrobial stewardship is a coordinated program that promotes the appropriate use of antimicrobials (including antibiotics), improves patient outcomes, reduces microbial resistance, and decreases the spread of infections caused by multidrug-resistant organisms.
WHO launched the Global Antimicrobial Resistance and Use Surveillance System (GLASS) in 2015 to fill knowledge gaps and inform strategies at all levels.
ACCORDING TO apic.org,
Antimicrobial stewardship is a coordinated program that promotes the appropriate use of antimicrobials (including antibiotics), improves patient outcomes, reduces microbial resistance, and decreases the spread of infections caused by multidrug-resistant organisms.
ACCORDING TO pewtrusts.org,
Antibiotic stewardship refers to efforts in doctors’ offices, hospitals, long term care facilities, and other health care settings to ensure that antibiotics are used only when necessary and appropriate
According to WHO,
Antimicrobial stewardship is a systematic approach to educate and support health care professionals to follow evidence-based guidelines for prescribing and administering antimicrobials
In 1996, John McGowan and Dale Gerding first applied the term antimicrobial stewardship, where they suggested a causal association between antimicrobial agent use and resistance. They also focused on the urgency of large-scale controlled trials of antimicrobial-use regulation employing sophisticated epidemiologic methods, molecular typing, and precise resistance mechanism analysis.
Antimicrobial Stewardship(AMS) refers to the optimal selection, dosing, and duration of antimicrobial treatment resulting in the best clinical outcome with minimal side effects to the patients and minimal impact on subsequent resistance.
According to the 2019 report, in the US, more than 2.8 million antibiotic-resistant infections occur each year, and more than 35000 people die. In addition to this, it also mentioned that 223,900 cases of Clostridoides difficile occurred in 2017, of which 12800 people died. The report did not include viruses or parasites
VISION
Being proactive
Supporting optimal animal and human health
Exploring ways to reduce overall use of antimicrobials
Using the drugs that prevent and treat disease by killing microscopic organisms in a responsible way
GOAL
to prevent the generation and spread of antimicrobial resistance (AMR). Doing so will preserve the effectiveness of these drugs in animals and humans for years to come.
being to preserve human and animal health and the effectiveness of antimicrobial medications.
to implement a multidisciplinary approach in assembling a stewardship team to include an infectious disease physician, a clinical pharmacist with infectious diseases training, infection preventionist, and a close collaboration with the staff in the clinical microbiology laboratory
to prevent antimicrobial overuse, misuse and abuse.
to minimize the developme
Delivering More Efficient Therapeutic Protein Expression Systems Through Cell Line Engineering Applications
1. Trissa Borgschulte, PhD
Head of Cell Line Development and Engineering
Process Solutions Upstream R&D
Applications in Therapeutic Protein Expression Systems
NOT FOR U.S. AND CANADA AUDIENCE
3. 3
Industry feedback
Webinar: Cell Line Engineering
Support the use of
concentrated feeds
Provide for reduced
analytical workload
Support clone stability
Maintain/enable favorable PQ
attributes
Reduce regulatory
compliance costs
Control/reduce metabolic
waste
Reduce product and process
related impurities
Reduce regulatory and risk
considerations
Increase process robustness
Promote high product yield
Support high growth rates
and cell densities
Scalable process
Provide batch to batch
consistency
Support single use
disposables
Downstream process friendly
Promote culture longevity
Maintain product stability
through harvest
Reduce product
heterogeneity
What makes a robust
therapeutic protein
manufacturing
process?
4. 4
Webinar: Cell Line Engineering
Cell Line Development and Engineering Strategy
Sources of Cell Line Engineering Targets
Discovery
Literature
Customer requests
Applications in Biomanufacturing
Selection mechanisms
Cell culture performance
Anti-apoptotic, metabolic engineering
Glycoengineering
Host cell proteins
Risk mitigation
Viral resistance
Targeted gene integration
Understanding
CHO Cell
Biology
Cell Line
Engineering
Media, Feed
and Process
Optimization
5. 5 Webinar: Cell Line Engineering
Trait Stacking to Improve Therapeutic Protein Manufacturing
Processes and Products
TI
Streamlined CLD Process and Enhanced
Performance
Homogeneity, stability
More Effective Biologics
Removal of immunogenic sugars, increased drug half-life
Robust CLD Process and High Performing
Recombinant Clones
Reduced time lines, high titers, increased stability
More Efficient Manufacturing Processes
Viral resistant cell lines, removal of contaminating host cell
proteins
Strong Base
Suspension adapted, CD media, bioreactor robust
Glyco-
engineering
Risk
Mitigation
CHO GS-/-
CHO K1
6. 6 Webinar: Cell Line Engineering
Genome Editing with ZFNs
Zinc Finger Nucleases
Introduce double strand breaks at sequence
specific genomic loci
Limited off-target effects
Cellular DNA Repair Mechanisms
NHEJ – imperfect repair resulting in INDELS
(gene KO)
HR – “perfect” repair by copying off of a template
(targeted integration)
Cellular
DNA
Repair
DNA
Binding
and
Cutting
7. 7 Webinar: Cell Line Engineering
Cell Line Engineering Workflow
ZFN Transfection
Activity Confirmation
(Cel 1 assay)
Single Cell Cloning
Expansion and
Further
Characterization
PCR and Seq Analysis
Phenotypic Assay
(if available)
8. 8 Webinar: Cell Line Engineering
Metabolic Selection: Benefits of a GS-/- Host Cell Line
GS-/- host cell lines lead to decreased CLD timelines and resources and
enhanced manufacturing performance
• Multiple rounds of
amplification are not
required
• Fewer clones evaluated
to identify high
performers
• Decreased CLD timelines
and resources
GS vs. DHFR
• MSX not required
• High performance in CD
processes
• Increased clone stability
• Fewer clones evaluated
to identify high
performers
• Decreased CLD timelines
and resources
GS-/- vs. GS+/+
• System saved 8 weeks
of development time
• Stable pool titers
exceeding 1g/L
• Clone titers averaging 2-
4.5 g/L
• Significant increase in
clone stability
• Fewer clones evaluated
to identify high
performers
GS-/- Industry
Feedback
9. 9 Webinar: Cell Line Engineering
CHO K1 GS-/- Cell Line Generation and Characterization
Genotype
Confirmation
•Genome sequencing
•qRT-PCR
•Western Blot
•Glutamine sensitivity
Phenotype
Characterization
•Robust growth/scalability in CD media
•Metabolic Profiling
•Transfectability
•Transient r-protein expression
•Cloning efficiencies
Performance
Validation
•Stable r-protein expression
•High pool and clone titers
•Long term stability
•Complex N-glycans
20 clones
3 clones
10. Transfection
-Gln Stable Pool Selection
(minipools)
Stable Pool Expansion and
Characterization
Single Cell Cloning of Lead Pools
Clone Expansion and
Characterization
10 Webinar: Cell Line Engineering
GS-/- Recombinant Cell Line Generation
7–9 weeks
7–9 weeks
11. 11 Webinar: Cell Line Engineering
GS-/- IgG Producing Stable Pools and Clones: Productivity Performance
Fed Batch TPP
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
Stable Pools -
Fed Batch
Clones - Fed
Batch
g/L
Titer Range of Top Expressing
GS-/- Pools and Clones
12. 12 Webinar: Cell Line Engineering
GS-/- IgG Stable Clones: Stability Performance
8 out of 10 top clones maintain >70% titer over 60 generations
13. 13 Webinar: Cell Line Engineering
Glycoengineering for Enhanced Product Quality
14. 14 Webinar: Cell Line Engineering
N-Glycan Biosynthesis in CHO
Modified from: Hossler et al., (2009) Glycobiology
GGTA
15. 15 Webinar: Cell Line Engineering
Engineering a Mgat1- Cell Line for Recombinant Proteins with High
Man5 Glycoforms
Sources:
1. Betting, et al. (2009) Vaccine
2. Lam, et al. (2005) J of Immunology
Targeting applications for therapeutic
proteins
Cerezyme® targets the mannose receptor on
macrophages
Carbohydrate remodeling required
Increased efficacy of mannose receptor
targeted vaccines
• Insect cell derived antigens are more effective for
targeting mannose receptors on APCs1
• Antigens produced in Pichia Pastoris are more
potent at inducing CD4+ T cell proliferation2
X-ray crystallography
• Increased homogeneity in protein structure
16. 16 Webinar: Cell Line Engineering
Mgat1- Cell Line Engineering in a Recombinant IgG Producing CHO
Clone
Lectin Enrichment of ZFN Transfected Pools
Ricinus communis agglutinin-I (RCA-I)
Lectin isolated from castor bean seeds
Binds to terminal galactose residues
Toxic to CHO cells with WT glycan profiles
Ladder Mock
ZFN
DNA
ZFN
RNA
ZFN
RNA
ZFN
DNA
Percent Modified 0 3.8 5.5 35.7 33.2
-RCA +RCA
17. 17 Webinar: Cell Line Engineering
Man5 Glycoform is the Predominant Species in Lectin Enriched Pools
Mock-transfected
RCA-I enriched
18. 18 Webinar: Cell Line Engineering
Man5 Glycoform is the Predominant Species in Lectin Enriched Pools
0
10
20
30
40
50
60
70
80
90
100
PercentGlycanSpecies
Mock
Mgat1 ZFN DNA
Mgat1 ZFN RNA
Mgat1 ZFN DNA + RCA-I
Mgat1 ZFN RNA + RCA-I
19. 19 Webinar: Cell Line Engineering
Mgat1- Clones Have Similar Growth and Productivity to WT Clones
0
1
2
3
4
5
6
7
8
9
0 2 4 6 8 10 12 14
VCD(10e6cells/ml)
Days in Culture
Original
WT1
WT2
KO1 -RCA
KO2 -RCA
KO3 +RCA
KO4 +RCA
KO5 +RCA
KO6 +RCA
0
200
400
600
800
1000
Original WT1 WT2 KO1 -
RCA
KO2 -
RCA
KO3
+RCA
KO4
+RCA
KO5
+RCA
KO6
+RCA
IgG(mg/L)
20. 20 Webinar: Cell Line Engineering
Mgat1- Clones Have Man5 as Predominant Glycan Species
0.0
10.0
20.0
30.0
40.0
50.0
60.0
70.0
80.0
90.0
PercentGlycanSpecies
Original
WT1
WT2
KO1 -RCA
KO2 -RCA
KO3 +RCA
KO4 +RCA
KO5 +RCA
KO6 +RCA
21. 21 Webinar: Cell Line Engineering
Mgat1- Cell Line Engineering in a CHO K1 GS-/- Host Cell Line:
Growth Performance
0
2
4
6
8
10
12
0 2 4 6 8 10 12
VCD(10e6cells/ml)
Days in Culture
WT
AB4
BC9
BD7
BH11
CG10
23. 23 Webinar: Cell Line Engineering
Man5 Glycoform is the Predominant Species in GS-/-/Mgat1- IgG Stable
Pools
GS-/-
IgG Stable Pool 1
GS-/-/Mgat1-
IgG Stable Pool 1
GS-/-/Mgat1-
IgG Stable Pool 2
24. 24 Webinar: Cell Line Engineering
Engineering CMAH-/- and GGTA-/- Cell Lines for Decreased
Biotherapeutic Immunogenicity
CMAH
CMP-Neu5Ac hydroxylase
Conversion of Neu5Ac to Neu5Gc sialic acid
N-acetylneuraminic acid (NANA or Neu5Ac)
N-glycolylneuraminic acid (NGNA or Neu5Gc)
Neu5Gc is not expressed in humans and can be
recognized as a foreign epitope
GGTA
N-acetyllactosaminide 3-α-galactosyltransferase-1
Immunogenic α-gal moiety
Expressed in murine cells but not in human
Humans have circulating antibodies to alpha-gal
Some CHO cell lines express functional Ggta1
25. 25 Webinar: Cell Line Engineering
CMAH-/-/GGTA-/- Cell Line Engineering in a CHO K1 GS-/- Host Cell
Line
GS-/- Host
CMAH ZFN Transfection
SCC and Genotyping
5 KO Clones
Clone Expansion and Characterization
Clone Banking
GS-/-/CMAH-/- Host
GGTA ZFN Transfection
SCC and Genotyping
1 KO Clone
Clone Expansion and Characterization
Clone Banking
26. 26 Webinar: Cell Line Engineering
GS-/-/CMAH-/-/GGTA-/- Host Cell Line Growth Characterization
0
1
2
3
4
5
6
7
8
9
10
0 2 4 6 8 10
VCD(10e6cells/ml)
Days in Culture
0.0
10.0
20.0
30.0
40.0
50.0
60.0
70.0
80.0
90.0
100.0
0 2 4 6 8
PercentViability
Days in Culture
28. 28 Webinar: Cell Line Engineering
Targeted Gene Integration for Biomanufacturing
Reduced variability/enhanced cell line
performance
• Decreased integration site side effects
• Clone to clone consistency
• Molecule to molecule consistency
Decreased cell line development timelines
• More homogeneous stable pools
• Decreased clone screening
Increased pool and clonal stability
• Use of well characterized safe harbor sites
• Remove stability from the critical path
29. 29 Webinar: Cell Line Engineering
Targeted Gene Integration Project Focus Areas
321
• Short RFLP donors
• GFP reporters
• IgG and Fc r-proteins
• Decreasing NHEJ
• Increasing HR
• Tagged ZFNs
• Optimized donor designs
• TI landing pads
• Reverse engineering of
high expressing clones
• Transciptomis analysis of
constitutively expressed
genes
Increased EfficiencyFeasibility Hot Spot Identification
30. 30 Webinar: Cell Line Engineering
Targeted Integration of GFP at the Actin Locus
DonorOnlyZFN+Donor
1.2% GFP+
2.4% GFP+
decreased scaling
actual
GFP Reporter HAHA
Actin GeneGFP Reporter Actin Gene
31. 31 Webinar: Cell Line Engineering
Targeted Integration of GFP at the Actin Locus
Mock
DonorOnly
Donor+ZFN-Lsort
Donor+ZFN-HsortGFP Reporter Actin Gene
32. 32 Webinar: Cell Line Engineering
Targeted Integration on IgG at a Safe Harbor Site
Transfection
• CHO SH1 ZFN RNA
• IgG/GS Plasmid Donor
Stable Pool
Generation
• GS Selection
• FACS Enrichment
SCC
Generation
• jPCR/Seq, qPCR
• G&P
• Stability
Targeted Integration SCC
Random Integration SCC
33. 33 Webinar: Cell Line Engineering
Summary
4
3
2
1
Genome editing is a valuable tool for enhancing biomanufacturing
expression systems
GS-/- host cell lines can lead to more efficient cell line development
processes and higher performing recombinant pools and clones
Glycoengineering can lead to more efficient manufacturing processes
and the production of more effective therapeutic proteins
Targeted integration can result in increased homogeneity and stability
of recombinant stable pools and clones
5
Full extent of cell line engineering applications in biomanufacturing
remains to be discovered
35. Erika Holroyd
Product Manager, Cell Line
Development and Engineering
CHOZN® Program
erika.holroyd@sial.com
CONTACT
Trissa Borgschulte, PhD
Head of Cell Line Development and Engineering
CHOZN® Program | Process Solutions Upstream R&D
trissa.borgschulte@sial.com
Contact