In this webinar, you will learn:
Intensified plasma Immunoglobulin purifications
Scalable process development with latest technologies
Improved safety and quality of plasma IgG meeting required quality attributes
Detailed description:
Plasma-derived immunoglobulins (IgG) are essential medicines that are in worldwide shortage. How to develop optimized processing steps for robust and efficient manufacturing has been a constant goal, to make the most out of the precious plasma raw material.
In this study, we present a worse-case equivalent of plasma intermediate, explore various process steps along the fractionation flow, including flow-through-mode chromatography, affinity chromatography, virus inactivation steps and removal of solvent/detergent, single-pass TFF (SPTFF), clarification, and aseptic filtration, to establish a robust, easy-to-operate, readily scalable plasma IgG process with over 99% purity, depletion of IgA, isoagglutinin, and thrombogenic markers, meeting the commonly required 20% concentration for subcutaneous IgG infusion. Such solutions would be appropriate for various IgG intermediates which help to improve the global supply of immunoglobulins.
Endotoxin Control and Clearance in BiomanufacturingMilliporeSigma
In this webinar, you will learn:
Sources of endotoxin contamination
Contamination control strategy
Endotoxin removal strategies
Detailed description:
Endotoxin, a lipopolysaccharide (LPS), is a type of pyrogen and is a component of the exterior cell wall of Gram-negative bacteria. To ensure safety on patient’s endotoxin content in the drug should always be controlled. In a biological processing it may emanate from facility, utility, raw materials, process, and personnel. In this webinar we discuss the regulatory norms, strategies for prevention & removal of endotoxin to ensure that the final drug product is safe.
Breaking the Status Quo: Using Mass Spectrometry to detect Host Cell ProteinsMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3b3Tbcd
Measurement of host cell proteins is vital to ensuring a biotherapy's purity and a patient's safety. Biotherapies treat diseases with products produced by living organisms, as a result, host cell components must be characterized and controlled. We'll review new methods within product characterization for detection.
Trace amounts of host cell proteins can be present after the production and purification of any biopharmaceutical. Detection of these species requires highly specific techniques to accurately quantify even low levels of contamination. Host cell protein impurities, present at PPM-levels in biotherapies, are a major immunogenicity risk because they can elicit an unpredictable immune response in patients. Their complex and diverse nature makes them challenging to detect or monitor. With acceptance criteria for host residual DNA usually set at a very low level (often =1.0 pg of DNA per mg of drug substance), effective removal techniques and sensitive methods of detection are critical.
Antibody-based techniques, like the enzyme-linked immunosorbent assay (ELISA), have been used to assess the HCP load of biotherapeutics before and after process changes. However, these techniques do not necessarily detect qualitative changes in the HCP population. In this webinar, we will discuss how mass spectrometry (MS)-based approaches coupled with ELISA methods help detect qualitative and quantitative differences in HCP populations.
In this webinar, you will learn:
• Comprehensive HCP ID and semi-quantitation
• HC agnostic process
• Creation of process specific database
• Differential clearance of specific HCPs throughout purification steps
• Monitoring of problematic species e.g. immunogenic (PLBL2), lipases and proteases
• Explanation about why 90% of BLAs filed included this HCP MS data
Latest Updates in Biosafety Testing for Gene TherapyMilliporeSigma
The field of Gene Therapy is moving at a fast pace providing promise of lifesaving medicines to previously unmet clinical needs. Of significant importance in the development of these novel therapies is the ability to demonstrate their safety including freedom from adventitious agents originating from raw materials or introduced during the manufacturing process.
It can be challenging, in such a fast moving field, to identify and navigate the relevant regulatory requirements and expectations for biosafety testing of such therapies. So too it can be difficult to select the optimal test methods in light of limited product availability and shelf life. Encompassing current biosafety testing approaches for bacteria, fungi, mycoplasma and viruses on starting materials to drug product, this webinar will provide you with the fundamentals to design your own Gene Therapy testing strategy.
In this webinar, you will learn:
• The most up to date regulatory expectations for Gene Therapies
• How to design a testing strategy to meet US FDA and EMA requirements
• How selecting the right biosafety test can overcome some of the unique challenges with Gene Therapies
Setting up for successful lot release testing by Edmund AngMilliporeSigma
Is your lot release testing strategy ready for global commercialization?
In this webinar, you will learn:
• CMC testing requirements with CHO production platform for global commercialization
• Lot release testing of product intermediates and final product
• Product-specific qualification study
• Alternative rapid testing methods to advance lot release testing
CHO cells continue to serve as a key cell substrate for the manufacturing of recombinant proteins that span beyond therapeutic monoclonal antibodies and including subunit vaccines.
In this presentation, we will cover the CMC testing requirements with CHO production platform for global commercialization, Lot release testing of product intermediates and final product, product-specific qualification study and highlight the application of new testing methods and the benefits they bring to advance Lot Release Testing.
Excipients selection for high risk formulations Smita RajputMerck Life Sciences
Are you choosing the right excipients for your high risk application? Find out how to select the right excipients and enable your process optimization to improve the total cost of ownership.
In this webinar, you will learn:
• Selection of right excipients for high risk formulation is very critical step
• Low Endotoxin and low bioburden limits are important aspect while selecting raw materials
• Strong regulatory support is crucial for high risk formulation
Excipients selection for high risk formulations like parenteral and ophthalmic applications is very challenging. Excipients should be inert with high purity for such dosage forms because trace amounts of impurities present in excipients can interact with active pharmaceutical ingredient (API) which results in instability of the formulation. This presentation discusses how to select the right excipients for high-risk applications and gives guidance for process optimization by choosing the best combination of filters and excipients to improve the total cost of ownership.
A Turn-Key Flow-Through-Mode Purification Process to improve Quality and Safe...Merck Life Sciences
In this webinar, you will learn:
Intensified plasma Immunoglobulin purifications
Scalable process development with latest technologies
Improved safety and quality of plasma IgG meeting required quality attributes
Detailed description:
Plasma-derived immunoglobulins (IgG) are essential medicines that are in worldwide shortage. How to develop optimized processing steps for robust and efficient manufacturing has been a constant goal, to make the most out of the precious plasma raw material.
In this study, we present a worse-case equivalent of plasma intermediate, explore various process steps along the fractionation flow, including flow-through-mode chromatography, affinity chromatography, virus inactivation steps and removal of solvent/detergent, single-pass TFF (SPTFF), clarification, and aseptic filtration, to establish a robust, easy-to-operate, readily scalable plasma IgG process with over 99% purity, depletion of IgA, isoagglutinin, and thrombogenic markers, meeting the commonly required 20% concentration for subcutaneous IgG infusion. Such solutions would be appropriate for various IgG intermediates which help to improve the global supply of immunoglobulins.
Looking for insights into current global regulatory expectations for viral safety? Read the special report from BioProcess International, in collaboration with Martin Wisher, Senior Regulatory Consultant focusing on BioReliance biosafety® services.
Endotoxin Control and Clearance in BiomanufacturingMilliporeSigma
In this webinar, you will learn:
Sources of endotoxin contamination
Contamination control strategy
Endotoxin removal strategies
Detailed description:
Endotoxin, a lipopolysaccharide (LPS), is a type of pyrogen and is a component of the exterior cell wall of Gram-negative bacteria. To ensure safety on patient’s endotoxin content in the drug should always be controlled. In a biological processing it may emanate from facility, utility, raw materials, process, and personnel. In this webinar we discuss the regulatory norms, strategies for prevention & removal of endotoxin to ensure that the final drug product is safe.
Breaking the Status Quo: Using Mass Spectrometry to detect Host Cell ProteinsMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3b3Tbcd
Measurement of host cell proteins is vital to ensuring a biotherapy's purity and a patient's safety. Biotherapies treat diseases with products produced by living organisms, as a result, host cell components must be characterized and controlled. We'll review new methods within product characterization for detection.
Trace amounts of host cell proteins can be present after the production and purification of any biopharmaceutical. Detection of these species requires highly specific techniques to accurately quantify even low levels of contamination. Host cell protein impurities, present at PPM-levels in biotherapies, are a major immunogenicity risk because they can elicit an unpredictable immune response in patients. Their complex and diverse nature makes them challenging to detect or monitor. With acceptance criteria for host residual DNA usually set at a very low level (often =1.0 pg of DNA per mg of drug substance), effective removal techniques and sensitive methods of detection are critical.
Antibody-based techniques, like the enzyme-linked immunosorbent assay (ELISA), have been used to assess the HCP load of biotherapeutics before and after process changes. However, these techniques do not necessarily detect qualitative changes in the HCP population. In this webinar, we will discuss how mass spectrometry (MS)-based approaches coupled with ELISA methods help detect qualitative and quantitative differences in HCP populations.
In this webinar, you will learn:
• Comprehensive HCP ID and semi-quantitation
• HC agnostic process
• Creation of process specific database
• Differential clearance of specific HCPs throughout purification steps
• Monitoring of problematic species e.g. immunogenic (PLBL2), lipases and proteases
• Explanation about why 90% of BLAs filed included this HCP MS data
Latest Updates in Biosafety Testing for Gene TherapyMilliporeSigma
The field of Gene Therapy is moving at a fast pace providing promise of lifesaving medicines to previously unmet clinical needs. Of significant importance in the development of these novel therapies is the ability to demonstrate their safety including freedom from adventitious agents originating from raw materials or introduced during the manufacturing process.
It can be challenging, in such a fast moving field, to identify and navigate the relevant regulatory requirements and expectations for biosafety testing of such therapies. So too it can be difficult to select the optimal test methods in light of limited product availability and shelf life. Encompassing current biosafety testing approaches for bacteria, fungi, mycoplasma and viruses on starting materials to drug product, this webinar will provide you with the fundamentals to design your own Gene Therapy testing strategy.
In this webinar, you will learn:
• The most up to date regulatory expectations for Gene Therapies
• How to design a testing strategy to meet US FDA and EMA requirements
• How selecting the right biosafety test can overcome some of the unique challenges with Gene Therapies
Setting up for successful lot release testing by Edmund AngMilliporeSigma
Is your lot release testing strategy ready for global commercialization?
In this webinar, you will learn:
• CMC testing requirements with CHO production platform for global commercialization
• Lot release testing of product intermediates and final product
• Product-specific qualification study
• Alternative rapid testing methods to advance lot release testing
CHO cells continue to serve as a key cell substrate for the manufacturing of recombinant proteins that span beyond therapeutic monoclonal antibodies and including subunit vaccines.
In this presentation, we will cover the CMC testing requirements with CHO production platform for global commercialization, Lot release testing of product intermediates and final product, product-specific qualification study and highlight the application of new testing methods and the benefits they bring to advance Lot Release Testing.
Excipients selection for high risk formulations Smita RajputMerck Life Sciences
Are you choosing the right excipients for your high risk application? Find out how to select the right excipients and enable your process optimization to improve the total cost of ownership.
In this webinar, you will learn:
• Selection of right excipients for high risk formulation is very critical step
• Low Endotoxin and low bioburden limits are important aspect while selecting raw materials
• Strong regulatory support is crucial for high risk formulation
Excipients selection for high risk formulations like parenteral and ophthalmic applications is very challenging. Excipients should be inert with high purity for such dosage forms because trace amounts of impurities present in excipients can interact with active pharmaceutical ingredient (API) which results in instability of the formulation. This presentation discusses how to select the right excipients for high-risk applications and gives guidance for process optimization by choosing the best combination of filters and excipients to improve the total cost of ownership.
A Turn-Key Flow-Through-Mode Purification Process to improve Quality and Safe...Merck Life Sciences
In this webinar, you will learn:
Intensified plasma Immunoglobulin purifications
Scalable process development with latest technologies
Improved safety and quality of plasma IgG meeting required quality attributes
Detailed description:
Plasma-derived immunoglobulins (IgG) are essential medicines that are in worldwide shortage. How to develop optimized processing steps for robust and efficient manufacturing has been a constant goal, to make the most out of the precious plasma raw material.
In this study, we present a worse-case equivalent of plasma intermediate, explore various process steps along the fractionation flow, including flow-through-mode chromatography, affinity chromatography, virus inactivation steps and removal of solvent/detergent, single-pass TFF (SPTFF), clarification, and aseptic filtration, to establish a robust, easy-to-operate, readily scalable plasma IgG process with over 99% purity, depletion of IgA, isoagglutinin, and thrombogenic markers, meeting the commonly required 20% concentration for subcutaneous IgG infusion. Such solutions would be appropriate for various IgG intermediates which help to improve the global supply of immunoglobulins.
Looking for insights into current global regulatory expectations for viral safety? Read the special report from BioProcess International, in collaboration with Martin Wisher, Senior Regulatory Consultant focusing on BioReliance biosafety® services.
Keeping the (Adventitious) Virus Out of the (Adeno-Associated) VirusMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/2VRylbi
How can you keep an adventitious virus from contaminating your gene therapy that is delivered by an adeno virus vector? As viral vector bioprocessing advances, regulatory requirements for viral safety will as well. Learn how to define your viral clearance strategy for AAV delivered gene therapies.
How do you define a strategy for viral clearance for a process that inherently aims at purifying a virus?
Gene delivery using AAV has received a boost from two major approvals and the nearly 300 programs in the clinic. Novel gene therapies using viral vectors enable companies to transform the lives of people living with certain rare and ultra-rare diseases where treatments are often not available currently. Amongst a multitude of challenges in viral vector bioprocessing, uncertainty in regulatory expectations is a major challenge to gene therapy developers. Regulatory requirements are evolving as the science and manufacturing matures with more stringent measures for viral safety assurance expected for future approvals.
Learn how to implement techniques for adventitious virus removal in your viral vector process; we will focus on strategies for viral clearance along your journey towards commercial readiness of AAV-based processes.
In this webinar, you will learn:
• AAV process flows and focus areas for viral safety
• Strategies for implementing viral clearance measures in bioprocessing
• Case studies and data driven approaches on log reduction values (LRV) in a viral vector process
• Best practices and evaluation roadmaps on conducting viral clearance studies
Presented by: Ratish Krishnan, Senior Strategy Consultant, Novel Modalities Bioprocessing
Accelerating COVID-19 Therapies: How a streamlined biosafety strategy can get...Merck Life Sciences
Access the interactive recording: https://bit.ly/2xB2eRs
Abstract:
Vaccine and other biologic developers have long relied on traditional, growth-based methods for the detection of adventitious agents in a biosafety testing package. However, at a time where speed is of the essence, relying on testing methods that take many weeks is a real concern. Fortunately, alternative rapid detection methods can shorten timelines significantly — especially for Phase I testing. Here we will take you through these rapid alternatives and outline a testing strategy that can bring your therapy to the clinic faster.
Turning up the Compen-DIAL: Rapid Test Methods for Cell & Gene TherapiesMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3aeCPNB
Find out how we turn up the dial on quality control testing for cell and gene therapies through rapid methods for sterility, mycoplasma, and replication competent virus. We will review the current regulatory expectations as well as the benefits and limitations that come with each method.
Two of the biggest challenges with applying traditional quality control (QC) test methods to cell and gene therapies, is time to results, due to short shelf-life, and availability of sufficient sample, due to small production volumes.
So how can these challenges be overcome while still meeting regulatory expectations?
In this webinar we will discuss and review suitable methods for rapid testing of short-life cell and gene therapies that may also help conserve limited production material. We will look at benefits, limitations, and regulatory expectations for various QC needs including current and future rapid methods for sterility, mycoplasma and replication competent virus.
In this webinar, you will learn:
• Why the shelf life of a cell or gene therapy product may impact your QC testing strategy
• Current regulatory expectations surrounding rapid methods for sterility, mycoplasma and replication competent virus
• Potential impacts of pursuing a non-optimal QC testing strategy
Sartorius is a well respected global solution provider within the biologics industry, especially for antibody and vaccine production. Our proven products and services are being diversified for upstream and downstream processing of cells and viruses for allogeneic and autologous advanced therapies.
Sartorius provides the cellular immunotherapy industry with a range of scalable single-use production technologies. Our portfolio supports viral vector transduction, cell expansion, and downstream processing steps including harvest, wash, and concentration of cells
Webinar: Benefits of Monodisperse Activated PEGs in ADC DevelopmentMilliporeSigma
Watch the webinar here: http://bit.ly/PEGsWebinar
As the field of antibody-drug conjugation chemistry has advanced, the use of linkers to impart "drugability" has also been growing. Recent literature shows a variety of linear and branched monodisperse PEGs being called on to modify the PK/PD profile of some of the most active but lipophilic payloads. In this webinar, we will survey the types of PEGs that are used in ADCs and give examples of successful implementation to change the biophysical properties of the construct. In addition, we will focus on PEGs and why activated PEGs are widely used to improve the pharmacokinetics of drugs (such as pegylated proteins, peptides etc.). Raw materials used in this field may have a variety of polydispersity. However, when it comes to the use of activated PEGs as linkers for ADCs, the requirements are significantly higher. Therefore, the choice and control of the PEG linker is crucial for the successful development and accelerated time to market for your ADC.
This webinar addresses critical success factors such as:
• Survey of current PEG enhanced ADCs
• Why PEGs are used designing ADCs
• Critical parameters for aPEGs used as linkers
• Requirements in terms of analytical capabilities
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.
Innovation in Filter Validation and Technology TransferMilliporeSigma
Regulatory and manufacturing requirements exist to perform product-specific microbial retention testing on sterilizing filters. The implementation of a Quality by Design approach to microbial retention testing supports a paradigm that would obviate the need for product-specific testing for early stage products that do not have the quantity of material required to easily and efficiently perform such testing. Process and product parameters were varied to determine their effect on microbial retention.
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.emdmillipore.com/mlab
Welcome to the M Lab™ Collaboration Centers – where customers can use non-GMP lab spaces to operate equipment, evaluate processes and receive real-time technical support without disrupting production.
Plan your visit: www.merckmillipore.com/mlab
Upstream Viral Safety – Protect your bioreactor with Virus FiltrationMilliporeSigma
This poster summarizes the performance of a filter specifically developed for virus removal from chemically defined cell culture media. The filter removes high levels of virus, mycoplasma and bacteria without impacting cell growth, antibody titer, or protein quality. The filter has robust performance over a broad range of conditions offering an effective, easy to implement solution for media treatment.
Process Impurities: Don’t Let PEI or HCP Derail Your BioTherapyMerck Life Sciences
View our webinar here: https://bit.ly/2lKNdWX
Many different impurities are present in or generated during biotherapy manufacturing. This webinar will address how process contaminates can arise from raw input materials, occur as residual processing agents, or form as reaction by-products. We will review strategies within product characterization to de-risk the manufacturing process, including the use of routine and high complexity assays; and the recommended testing to meet regulatory requirements for clinical submission. Learn methods to avoid costly pitfalls and implement procedures to expedite product quality decisions at critical junctures in your development plan. We will discuss two types of therapies:
Cell & Gene Therapies
Polyethylenimine (PEI) is a transfection agent used in nearly all cell and gene therapy products. We will review the regulations and the liquid chromatography with charged aerosol detection (LC-CAD) methodology to demonstrate PEI removal during the production process.
Monoclonal Antibodies (mAb) and Cell & Gene Therapies
During mAb manufacturing and inherent to Cell & Gene Therapies, a significant proportion of process impurities arise from the host cell used to express the drug. Host cell protein (HCP) impurities, present at PPM-levels, are a major immunogenicity risk because they can elicit an unpredictable immune response in patients. We will review why their complex and diverse nature makes them challenging to monitor, and theho best practices, specifically HCP identification by mass spectrometry, for detection.
Learning points:
1. Accurate detection and characterization of residual PEI in cell and gene therapy products
2. Effective detection and characterization of residual host cell proteins (HCP) in mAbs
3. Available technology and assays for quantifying process impurities
4. Current regulatory requirements for detecting, quantifying, and removing process impurities during biotherapy manufacturing
Parvovirus Filtration Best Practices - 25 Years of Hands-On ExperienceMerck Life Sciences
In this webinar, you will learn:
- how to measure filter performance and capacity,
- how to optimize filter virus removal capability,
- and avoid potential pit-falls
Detailed description:
This webinar will cover all aspects of parvovirus filtration best practices: process development/ optimization, pilot scale-up, and validation and explain the important connections between these activities. The rationale for the recommended best practices will be explained by discussing the underlying mechanisms that control filter performance.
Long Acting Injectables - A New Dimension for Proteins and PeptidesMerck Life Sciences
Access the recording: https://bit.ly/2xAaMba
Abstract:
Long acting injectables (LAI) have been around for decades for the delivery of small molecules and peptides to treat chronic and site-specific diseases. However, when it comes to more sensitive biological therapeutics the classical polylactide and polylactide/glycolide based systems suffer from several limitations (e.g. uncontrolled release kinetics, in situ pH drop, protein degradation) making them unsuitable. The SynBiosys® biodegradable polymeric microparticle technology combines all the features required for LAI formulations for biologics. In two case studies we will showcase sustained release formulations for peptides and proteins and demonstrate their potential via extensive in vitro and in vivo characterization.
See the Whole Picture: Using SV-AUC for Empty/Full AAV Capsid AnalysisMilliporeSigma
Watch this webinar here: https://bit.ly/31ZZM3n
Join this webinar for key insights on using the SV-AUC assay for empty/full analysis of your AAV viral vector. We’ll cover the technical requirements for this assay, data interpretation, and finally how this assay fits into the larger picture of AAV characterization.
Recombinant adeno-associated viruses (AAV) are widely used as gene transfer vectors. However, AAV production generates mixed populations of viral capsids containing either complete viral vector genome (full capsids); partially filled, and those lacking the viral genome (empty capsids). Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC) offers a robust, accurate, and consistent method for characterizing empty/full AAV capsid composition. In this webinar we will review the key technical requirements for performing an AUC assay as well as analysis and data interpretation of the results generated.
In this webinar, you will learn:
• Regulatory expectations for empty/full analysis
• Key technical requirements for running an AUC assay and how to interpret the data from the results generated
• How the AUC assay fits into the larger picture of AAV characterization
Welcome to the M Lab™ Collaboration Centers – where customers can use non-GMP lab spaces to operate equipment, evaluate processes and receive real-time technical support without disrupting production.
Plan your visit: www.merckmillipore.com/mlab
Upcoming USP 665 - Level of Characterization of Single-Use Systems Today and ...MilliporeSigma
Register for the interactive, on-demand webinar now: https://bit.ly/USP665
Single-use plastic systems are being utilized more frequently especially for COVID-19 vaccine manufacturing. However, there are issues regarding standardization of quality information that limits implementation efficiencies. One of the challenges is the evaluation of leachables derived from a variety of different plastic components in a timely manner.
Since the USP <665> highlights a risk assessment approach with no typical pass/fail limit, approaches to decision-making based on the extractables data package will be reviewed. In addition, we will highlight legacy testing requirements which may not be necessary once USP <665> is implemented.
In this webinar, we will discuss:
- Regulatory expectations of extractables and leachables assessment today and tomorrow
- The right criteria that need to be assessed to select the type and quality of plastic materials for use in biopharmaceutical manufacturing
Chromatography: Chromatographic strategies for IVIG purification – Part 2MilliporeSigma
Immunoglobulin G (IgG) is an important plasma-derived product used to treat patients with primary immunodeficiency or auto-immune diseases. Industrial plasma fractionators are increasingly using chromatography as an essential step of the purification of IgG for therapeutic use. Therapeutic IgG should meet quality criteria including a low residual level of contamination with other proteins such as IgA, IgM, proteolytic enzymes, or Factor XI/XIa. We have developed a chromatographic method using our commercially available Fractogel® EMD TMAE(M) resin that allows the purification of plasma-derived IgG in a negative mode, whereby contaminants such as IgA and IgM are adsorbed on the resin and further eliminated by subsequent elution prior to column regeneration and re-use for the next cycle. In this study we show a systematic approach in chromatographic process development to determine the optimal purification conditions for the plasma-derived IgG based on 5%-pH 5.5 caprylic acid treatment (CA-IgG). The reliability and consistency of the resin in IgG purification was evaluated up to 200 chromatographic cycles.
Results show that under worst-case conditions of a crude IgG feed with high linear flow rate, there is powerful removal of both IgA and IgM from the IgG flow-through corresponding to 75-90 mg of proteins/mL of resin. The robustness of Fractogel® EMD TMAE(M) resin at pH 6.0 was confirmed over 10 cycle runs, and the wash fraction was proportionally enriched in IgA and IgM, confirming the adsorption on the resin. The recovery in IgG subclasses was good, and improvement of IgG purity from around 82% to around 97% purity was achieved. Furthermore, there was no generation of thrombogenicity during the chromatography based on several assays including TGA. The optimal chromatographic conditions/parameters were confirmed with the use of a controlled IgG with a loading close to 199 mg of IgG/ml of resin. Finally, in the 200-cycle study, Fractogel® EMD TMAE(M) resin demonstrated excellent resistance for over 201 cycles in its capacity to reproducibly remove IgA and IgM from a crude CA-IgG preparation, showed good recovery of IgG without affecting the IgG sub-class distribution, and even suggested a potential chromatographic removal of thrombogenic factors developing in the CA-IgG feed over time.
In this webinar, you will learn:
- How to systematically set up an IVIG purification strategy.
- How to evaluate the quality of purified IVIG.
- How to confirm the robustness of an upgraded IVIG purification process.
Chromatography: Chromatographic strategies for IVIG purification – Part 2Merck Life Sciences
Immunoglobulin G (IgG) is an important plasma-derived product used to treat patients with primary immunodeficiency or auto-immune diseases. Industrial plasma fractionators are increasingly using chromatography as an essential step of the purification of IgG for therapeutic use. Therapeutic IgG should meet quality criteria including a low residual level of contamination with other proteins such as IgA, IgM, proteolytic enzymes, or Factor XI/XIa. We have developed a chromatographic method using our commercially available Fractogel® EMD TMAE(M) resin that allows the purification of plasma-derived IgG in a negative mode, whereby contaminants such as IgA and IgM are adsorbed on the resin and further eliminated by subsequent elution prior to column regeneration and re-use for the next cycle. In this study we show a systematic approach in chromatographic process development to determine the optimal purification conditions for the plasma-derived IgG based on 5%-pH 5.5 caprylic acid treatment (CA-IgG). The reliability and consistency of the resin in IgG purification was evaluated up to 200 chromatographic cycles.
Results show that under worst-case conditions of a crude IgG feed with high linear flow rate, there is powerful removal of both IgA and IgM from the IgG flow-through corresponding to 75-90 mg of proteins/mL of resin. The robustness of Fractogel® EMD TMAE(M) resin at pH 6.0 was confirmed over 10 cycle runs, and the wash fraction was proportionally enriched in IgA and IgM, confirming the adsorption on the resin. The recovery in IgG subclasses was good, and improvement of IgG purity from around 82% to around 97% purity was achieved. Furthermore, there was no generation of thrombogenicity during the chromatography based on several assays including TGA. The optimal chromatographic conditions/parameters were confirmed with the use of a controlled IgG with a loading close to 199 mg of IgG/ml of resin. Finally, in the 200-cycle study, Fractogel® EMD TMAE(M) resin demonstrated excellent resistance for over 201 cycles in its capacity to reproducibly remove IgA and IgM from a crude CA-IgG preparation, showed good recovery of IgG without affecting the IgG sub-class distribution, and even suggested a potential chromatographic removal of thrombogenic factors developing in the CA-IgG feed over time.
In this webinar, you will learn:
- How to systematically set up an IVIG purification strategy.
- How to evaluate the quality of purified IVIG.
- How to confirm the robustness of an upgraded IVIG purification process.
This presentation reviews current trends in bioprocessing purification and includes key considerations for continuous processing and connected polishing for monoclonal antibodies. Topics include:
• Market trends and the evolution of next-generation processes
• Intensified capture processing
• Continuous virus inactivation
• Connected flow-through polishing
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/mlab
Technology Trends in Bioprocessing PurificationMilliporeSigma
This presentation reviews current trends in bioprocessing purification and includes key considerations for continuous processing and connected polishing for monoclonal antibodies. Topics include:
• Market trends and the evolution of next-generation processes
• Intensified capture processing
• Continuous virus inactivation
• Connected flow-through polishing
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.emdmillipore.com/mlab
Keeping the (Adventitious) Virus Out of the (Adeno-Associated) VirusMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/2VRylbi
How can you keep an adventitious virus from contaminating your gene therapy that is delivered by an adeno virus vector? As viral vector bioprocessing advances, regulatory requirements for viral safety will as well. Learn how to define your viral clearance strategy for AAV delivered gene therapies.
How do you define a strategy for viral clearance for a process that inherently aims at purifying a virus?
Gene delivery using AAV has received a boost from two major approvals and the nearly 300 programs in the clinic. Novel gene therapies using viral vectors enable companies to transform the lives of people living with certain rare and ultra-rare diseases where treatments are often not available currently. Amongst a multitude of challenges in viral vector bioprocessing, uncertainty in regulatory expectations is a major challenge to gene therapy developers. Regulatory requirements are evolving as the science and manufacturing matures with more stringent measures for viral safety assurance expected for future approvals.
Learn how to implement techniques for adventitious virus removal in your viral vector process; we will focus on strategies for viral clearance along your journey towards commercial readiness of AAV-based processes.
In this webinar, you will learn:
• AAV process flows and focus areas for viral safety
• Strategies for implementing viral clearance measures in bioprocessing
• Case studies and data driven approaches on log reduction values (LRV) in a viral vector process
• Best practices and evaluation roadmaps on conducting viral clearance studies
Presented by: Ratish Krishnan, Senior Strategy Consultant, Novel Modalities Bioprocessing
Accelerating COVID-19 Therapies: How a streamlined biosafety strategy can get...Merck Life Sciences
Access the interactive recording: https://bit.ly/2xB2eRs
Abstract:
Vaccine and other biologic developers have long relied on traditional, growth-based methods for the detection of adventitious agents in a biosafety testing package. However, at a time where speed is of the essence, relying on testing methods that take many weeks is a real concern. Fortunately, alternative rapid detection methods can shorten timelines significantly — especially for Phase I testing. Here we will take you through these rapid alternatives and outline a testing strategy that can bring your therapy to the clinic faster.
Turning up the Compen-DIAL: Rapid Test Methods for Cell & Gene TherapiesMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3aeCPNB
Find out how we turn up the dial on quality control testing for cell and gene therapies through rapid methods for sterility, mycoplasma, and replication competent virus. We will review the current regulatory expectations as well as the benefits and limitations that come with each method.
Two of the biggest challenges with applying traditional quality control (QC) test methods to cell and gene therapies, is time to results, due to short shelf-life, and availability of sufficient sample, due to small production volumes.
So how can these challenges be overcome while still meeting regulatory expectations?
In this webinar we will discuss and review suitable methods for rapid testing of short-life cell and gene therapies that may also help conserve limited production material. We will look at benefits, limitations, and regulatory expectations for various QC needs including current and future rapid methods for sterility, mycoplasma and replication competent virus.
In this webinar, you will learn:
• Why the shelf life of a cell or gene therapy product may impact your QC testing strategy
• Current regulatory expectations surrounding rapid methods for sterility, mycoplasma and replication competent virus
• Potential impacts of pursuing a non-optimal QC testing strategy
Sartorius is a well respected global solution provider within the biologics industry, especially for antibody and vaccine production. Our proven products and services are being diversified for upstream and downstream processing of cells and viruses for allogeneic and autologous advanced therapies.
Sartorius provides the cellular immunotherapy industry with a range of scalable single-use production technologies. Our portfolio supports viral vector transduction, cell expansion, and downstream processing steps including harvest, wash, and concentration of cells
Webinar: Benefits of Monodisperse Activated PEGs in ADC DevelopmentMilliporeSigma
Watch the webinar here: http://bit.ly/PEGsWebinar
As the field of antibody-drug conjugation chemistry has advanced, the use of linkers to impart "drugability" has also been growing. Recent literature shows a variety of linear and branched monodisperse PEGs being called on to modify the PK/PD profile of some of the most active but lipophilic payloads. In this webinar, we will survey the types of PEGs that are used in ADCs and give examples of successful implementation to change the biophysical properties of the construct. In addition, we will focus on PEGs and why activated PEGs are widely used to improve the pharmacokinetics of drugs (such as pegylated proteins, peptides etc.). Raw materials used in this field may have a variety of polydispersity. However, when it comes to the use of activated PEGs as linkers for ADCs, the requirements are significantly higher. Therefore, the choice and control of the PEG linker is crucial for the successful development and accelerated time to market for your ADC.
This webinar addresses critical success factors such as:
• Survey of current PEG enhanced ADCs
• Why PEGs are used designing ADCs
• Critical parameters for aPEGs used as linkers
• Requirements in terms of analytical capabilities
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.
Innovation in Filter Validation and Technology TransferMilliporeSigma
Regulatory and manufacturing requirements exist to perform product-specific microbial retention testing on sterilizing filters. The implementation of a Quality by Design approach to microbial retention testing supports a paradigm that would obviate the need for product-specific testing for early stage products that do not have the quantity of material required to easily and efficiently perform such testing. Process and product parameters were varied to determine their effect on microbial retention.
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.emdmillipore.com/mlab
Welcome to the M Lab™ Collaboration Centers – where customers can use non-GMP lab spaces to operate equipment, evaluate processes and receive real-time technical support without disrupting production.
Plan your visit: www.merckmillipore.com/mlab
Upstream Viral Safety – Protect your bioreactor with Virus FiltrationMilliporeSigma
This poster summarizes the performance of a filter specifically developed for virus removal from chemically defined cell culture media. The filter removes high levels of virus, mycoplasma and bacteria without impacting cell growth, antibody titer, or protein quality. The filter has robust performance over a broad range of conditions offering an effective, easy to implement solution for media treatment.
Process Impurities: Don’t Let PEI or HCP Derail Your BioTherapyMerck Life Sciences
View our webinar here: https://bit.ly/2lKNdWX
Many different impurities are present in or generated during biotherapy manufacturing. This webinar will address how process contaminates can arise from raw input materials, occur as residual processing agents, or form as reaction by-products. We will review strategies within product characterization to de-risk the manufacturing process, including the use of routine and high complexity assays; and the recommended testing to meet regulatory requirements for clinical submission. Learn methods to avoid costly pitfalls and implement procedures to expedite product quality decisions at critical junctures in your development plan. We will discuss two types of therapies:
Cell & Gene Therapies
Polyethylenimine (PEI) is a transfection agent used in nearly all cell and gene therapy products. We will review the regulations and the liquid chromatography with charged aerosol detection (LC-CAD) methodology to demonstrate PEI removal during the production process.
Monoclonal Antibodies (mAb) and Cell & Gene Therapies
During mAb manufacturing and inherent to Cell & Gene Therapies, a significant proportion of process impurities arise from the host cell used to express the drug. Host cell protein (HCP) impurities, present at PPM-levels, are a major immunogenicity risk because they can elicit an unpredictable immune response in patients. We will review why their complex and diverse nature makes them challenging to monitor, and theho best practices, specifically HCP identification by mass spectrometry, for detection.
Learning points:
1. Accurate detection and characterization of residual PEI in cell and gene therapy products
2. Effective detection and characterization of residual host cell proteins (HCP) in mAbs
3. Available technology and assays for quantifying process impurities
4. Current regulatory requirements for detecting, quantifying, and removing process impurities during biotherapy manufacturing
Parvovirus Filtration Best Practices - 25 Years of Hands-On ExperienceMerck Life Sciences
In this webinar, you will learn:
- how to measure filter performance and capacity,
- how to optimize filter virus removal capability,
- and avoid potential pit-falls
Detailed description:
This webinar will cover all aspects of parvovirus filtration best practices: process development/ optimization, pilot scale-up, and validation and explain the important connections between these activities. The rationale for the recommended best practices will be explained by discussing the underlying mechanisms that control filter performance.
Long Acting Injectables - A New Dimension for Proteins and PeptidesMerck Life Sciences
Access the recording: https://bit.ly/2xAaMba
Abstract:
Long acting injectables (LAI) have been around for decades for the delivery of small molecules and peptides to treat chronic and site-specific diseases. However, when it comes to more sensitive biological therapeutics the classical polylactide and polylactide/glycolide based systems suffer from several limitations (e.g. uncontrolled release kinetics, in situ pH drop, protein degradation) making them unsuitable. The SynBiosys® biodegradable polymeric microparticle technology combines all the features required for LAI formulations for biologics. In two case studies we will showcase sustained release formulations for peptides and proteins and demonstrate their potential via extensive in vitro and in vivo characterization.
See the Whole Picture: Using SV-AUC for Empty/Full AAV Capsid AnalysisMilliporeSigma
Watch this webinar here: https://bit.ly/31ZZM3n
Join this webinar for key insights on using the SV-AUC assay for empty/full analysis of your AAV viral vector. We’ll cover the technical requirements for this assay, data interpretation, and finally how this assay fits into the larger picture of AAV characterization.
Recombinant adeno-associated viruses (AAV) are widely used as gene transfer vectors. However, AAV production generates mixed populations of viral capsids containing either complete viral vector genome (full capsids); partially filled, and those lacking the viral genome (empty capsids). Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC) offers a robust, accurate, and consistent method for characterizing empty/full AAV capsid composition. In this webinar we will review the key technical requirements for performing an AUC assay as well as analysis and data interpretation of the results generated.
In this webinar, you will learn:
• Regulatory expectations for empty/full analysis
• Key technical requirements for running an AUC assay and how to interpret the data from the results generated
• How the AUC assay fits into the larger picture of AAV characterization
Welcome to the M Lab™ Collaboration Centers – where customers can use non-GMP lab spaces to operate equipment, evaluate processes and receive real-time technical support without disrupting production.
Plan your visit: www.merckmillipore.com/mlab
Upcoming USP 665 - Level of Characterization of Single-Use Systems Today and ...MilliporeSigma
Register for the interactive, on-demand webinar now: https://bit.ly/USP665
Single-use plastic systems are being utilized more frequently especially for COVID-19 vaccine manufacturing. However, there are issues regarding standardization of quality information that limits implementation efficiencies. One of the challenges is the evaluation of leachables derived from a variety of different plastic components in a timely manner.
Since the USP <665> highlights a risk assessment approach with no typical pass/fail limit, approaches to decision-making based on the extractables data package will be reviewed. In addition, we will highlight legacy testing requirements which may not be necessary once USP <665> is implemented.
In this webinar, we will discuss:
- Regulatory expectations of extractables and leachables assessment today and tomorrow
- The right criteria that need to be assessed to select the type and quality of plastic materials for use in biopharmaceutical manufacturing
Chromatography: Chromatographic strategies for IVIG purification – Part 2MilliporeSigma
Immunoglobulin G (IgG) is an important plasma-derived product used to treat patients with primary immunodeficiency or auto-immune diseases. Industrial plasma fractionators are increasingly using chromatography as an essential step of the purification of IgG for therapeutic use. Therapeutic IgG should meet quality criteria including a low residual level of contamination with other proteins such as IgA, IgM, proteolytic enzymes, or Factor XI/XIa. We have developed a chromatographic method using our commercially available Fractogel® EMD TMAE(M) resin that allows the purification of plasma-derived IgG in a negative mode, whereby contaminants such as IgA and IgM are adsorbed on the resin and further eliminated by subsequent elution prior to column regeneration and re-use for the next cycle. In this study we show a systematic approach in chromatographic process development to determine the optimal purification conditions for the plasma-derived IgG based on 5%-pH 5.5 caprylic acid treatment (CA-IgG). The reliability and consistency of the resin in IgG purification was evaluated up to 200 chromatographic cycles.
Results show that under worst-case conditions of a crude IgG feed with high linear flow rate, there is powerful removal of both IgA and IgM from the IgG flow-through corresponding to 75-90 mg of proteins/mL of resin. The robustness of Fractogel® EMD TMAE(M) resin at pH 6.0 was confirmed over 10 cycle runs, and the wash fraction was proportionally enriched in IgA and IgM, confirming the adsorption on the resin. The recovery in IgG subclasses was good, and improvement of IgG purity from around 82% to around 97% purity was achieved. Furthermore, there was no generation of thrombogenicity during the chromatography based on several assays including TGA. The optimal chromatographic conditions/parameters were confirmed with the use of a controlled IgG with a loading close to 199 mg of IgG/ml of resin. Finally, in the 200-cycle study, Fractogel® EMD TMAE(M) resin demonstrated excellent resistance for over 201 cycles in its capacity to reproducibly remove IgA and IgM from a crude CA-IgG preparation, showed good recovery of IgG without affecting the IgG sub-class distribution, and even suggested a potential chromatographic removal of thrombogenic factors developing in the CA-IgG feed over time.
In this webinar, you will learn:
- How to systematically set up an IVIG purification strategy.
- How to evaluate the quality of purified IVIG.
- How to confirm the robustness of an upgraded IVIG purification process.
Chromatography: Chromatographic strategies for IVIG purification – Part 2Merck Life Sciences
Immunoglobulin G (IgG) is an important plasma-derived product used to treat patients with primary immunodeficiency or auto-immune diseases. Industrial plasma fractionators are increasingly using chromatography as an essential step of the purification of IgG for therapeutic use. Therapeutic IgG should meet quality criteria including a low residual level of contamination with other proteins such as IgA, IgM, proteolytic enzymes, or Factor XI/XIa. We have developed a chromatographic method using our commercially available Fractogel® EMD TMAE(M) resin that allows the purification of plasma-derived IgG in a negative mode, whereby contaminants such as IgA and IgM are adsorbed on the resin and further eliminated by subsequent elution prior to column regeneration and re-use for the next cycle. In this study we show a systematic approach in chromatographic process development to determine the optimal purification conditions for the plasma-derived IgG based on 5%-pH 5.5 caprylic acid treatment (CA-IgG). The reliability and consistency of the resin in IgG purification was evaluated up to 200 chromatographic cycles.
Results show that under worst-case conditions of a crude IgG feed with high linear flow rate, there is powerful removal of both IgA and IgM from the IgG flow-through corresponding to 75-90 mg of proteins/mL of resin. The robustness of Fractogel® EMD TMAE(M) resin at pH 6.0 was confirmed over 10 cycle runs, and the wash fraction was proportionally enriched in IgA and IgM, confirming the adsorption on the resin. The recovery in IgG subclasses was good, and improvement of IgG purity from around 82% to around 97% purity was achieved. Furthermore, there was no generation of thrombogenicity during the chromatography based on several assays including TGA. The optimal chromatographic conditions/parameters were confirmed with the use of a controlled IgG with a loading close to 199 mg of IgG/ml of resin. Finally, in the 200-cycle study, Fractogel® EMD TMAE(M) resin demonstrated excellent resistance for over 201 cycles in its capacity to reproducibly remove IgA and IgM from a crude CA-IgG preparation, showed good recovery of IgG without affecting the IgG sub-class distribution, and even suggested a potential chromatographic removal of thrombogenic factors developing in the CA-IgG feed over time.
In this webinar, you will learn:
- How to systematically set up an IVIG purification strategy.
- How to evaluate the quality of purified IVIG.
- How to confirm the robustness of an upgraded IVIG purification process.
This presentation reviews current trends in bioprocessing purification and includes key considerations for continuous processing and connected polishing for monoclonal antibodies. Topics include:
• Market trends and the evolution of next-generation processes
• Intensified capture processing
• Continuous virus inactivation
• Connected flow-through polishing
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/mlab
Technology Trends in Bioprocessing PurificationMilliporeSigma
This presentation reviews current trends in bioprocessing purification and includes key considerations for continuous processing and connected polishing for monoclonal antibodies. Topics include:
• Market trends and the evolution of next-generation processes
• Intensified capture processing
• Continuous virus inactivation
• Connected flow-through polishing
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: www.emdmillipore.com/mlab
The present article deals with design of antibacterial and antifungal pH-responsive hydrogels based on Quaternary Ammonium Functionalized-Tragacanth Gum (QTG) biopolymer as drug delivery systems. The antimicrobial effects of the graft-copolymer hydrogels QTG/ polyacrylic acid (QTG-AA) and QTG/polyacrylamide (QTG-AM) were investigated against fi ve standard microorganisms including Candida albicans, Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa. The results of the in-vitro release of quercetin as a drug from the functionalized copolymer hydrogels exhibited dependence on the pH, immersion time, medium, and temperature. All copolymer hydrogels demonstrated antibacterial and antifungal properties and moreover, QTG-AM copolymers presented higher antimicrobial activity than QTG-AA copolymers.
Characterization of intact antibodies by pre-fractionation using gel electrop...Expedeon
Antibodies represent an important class of proteins due to their central role in the immune response. Moreover, there is an increasing interest in the use of recombinant antibodies as novel drug therapies.
Protein Purification and Analysis with PhyNexus and Caliper LifeSciencesChris Suh
PhyTip columns perform high throughput protein separation on Caliper liquid handling robotics and proteins analyzed by Caliper LabChip GXII microfluidic device
Benzonase® endonuclease: use and clearance with a TFF step in a cell culture-...Frédéric Sengler
Benzonase® endonuclease is a powerful tool for degrading all
forms of (deoxy)ribonucleic acids (DNA/RNA) in cell culture
harvests to base pair (BP) lengths under 8 units. It is effective
over a wide range of conditions including temperature, pH, and
varying concentrations of Mg2+, detergents, chelating agents,
and monovalent cations. It is often employed in the production
of viral vaccines, completely digesting DNA and RNA to improve
clearance and reduce solution viscosity.
Removing Benzonase® endonuclease, a 60 kD dimer, from the
process stream post-treatment may be achieved with Tangential
Flow Filtration (TFF) with the appropriate molecular weight
cut-off membrane (MWCO), typically 300 kD. This process has
heretofore not been well described in the literature, so the
present study fills this gap pairing Benzonase® endonuclease
treatment of a DNA-spiked inactivated flu virus cell culture
harvest with Pellicon® 2 cassettes, for clearance of the digested
DNA and remaining Benzonase® endonuclease by diafiltration.
The Viscosity Reduction Platform: Viscosity-reducing excipients for improveme...MilliporeSigma
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 PurificationMilliporeSigma
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...MilliporeSigma
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...MilliporeSigma
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...MilliporeSigma
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...MilliporeSigma
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
The Future of Pharma- and Biopharmaceutical AuditsMilliporeSigma
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...MilliporeSigma
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 therapiesMilliporeSigma
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...MilliporeSigma
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...MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3NDNIKe
Automated, fit-for-purpose tools are essential in CAR-T processing to support sustainable manufacturing of clinical and market-approved cell therapy products. This webinar will discuss how the ekko™ Acoustic Cell Processing System uses acoustic technology as a touchless approach to manipulate cells, enabling a modular tool across the CAR-T manufacturing workflow. Typical performance of templated ekko™ System processes for DMSO washout of leukapheresis material, low volume and high cell concentrate for electroporation preparation, and harvest of expanded T cells will be reviewed.
This webinar will also give an early glimpse at the ekko™ Select System for unmatched T cell selection.
In this webinar, you will:
• Uncover how the ekko™ System supports the broad industrialization of cell therapy, with particular focus on how to achieve low volume, high concentrate cell product for critical transduction and transfection steps
• Discover how ekko™ System for wash and concentrate processes throughout the cell therapy workflow achieve high cell recovery, viability, and effective residual removal
• Preview to ekko™ Select, our cell therapy selection platform, to achieve unmatched ease-of-use with direct processing from leukopaks reducing the need for preparation steps
Presented by:
Benjamin Ross-Johnsrud, Acoustic Technology Expert
Robert Scott, Mechanical Engineer III
How does the ICH Q5A revision impact viral safety strategies for biologics?MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3t7X9tg
How does the ICH Q5A revision impact viral safety strategies for biologics?
Biologics continue to grow at a fast pace. Manufactured using cell lines of human or animal origin, these are at risk of viral contamination making safety strategies critical. A comprehensive risk mitigation strategy using multiple orthogonal measures is a regulatory expectation. ICH Q5A, the globally-harmonized guideline outlines the expectations. ICH Q5A is currently being revised to address recent scientific advancements including novel therapeutic modalities, new manufacturing paradigms, updates in viral clearance applications, and alternate detection technologies. We’ll discuss the expected changes and potential impact on viral safety strategies with case studies and examples.
In this webinar, you will learn about:
• The Importance of virus testing in biologics products
• Regulatory landscape, expectations for the Q5A revision
• What's new and changing
• Examples of alternate testing schedules, impact on viral clearance
Presented by:
Manjula Aysola, Senior Regulatory Consultant
Alison Armstrong, PhD, Sr. Director, Technical and Scientific Solutions
Improve Operational Efficiency by Over 30% with Product, Process, & Systems A...MilliporeSigma
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...MilliporeSigma
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 SystemsMilliporeSigma
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 ...MilliporeSigma
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...MilliporeSigma
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...MilliporeSigma
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
How to Accelerate and Enhance ADC TherapiesMilliporeSigma
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.
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.
Explore our infographic on 'Essential Metrics for Palliative Care Management' which highlights key performance indicators crucial for enhancing the quality and efficiency of palliative care services.
This visual guide breaks down important metrics across four categories: Patient-Centered Metrics, Care Efficiency Metrics, Quality of Life Metrics, and Staff Metrics. Each section is designed to help healthcare professionals monitor and improve care delivery for patients facing serious illnesses. Understand how to implement these metrics in your palliative care practices for better outcomes and higher satisfaction levels.
Health Education on prevention of hypertensionRadhika kulvi
Hypertension is a chronic condition of concern due to its role in the causation of coronary heart diseases. Hypertension is a worldwide epidemic and important risk factor for coronary artery disease, stroke and renal diseases. Blood pressure is the force exerted by the blood against the walls of the blood vessels and is sufficient to maintain tissue perfusion during activity and rest. Hypertension is sustained elevation of BP. In adults, HTN exists when systolic blood pressure is equal to or greater than 140mmHg or diastolic BP is equal to or greater than 90mmHg. The
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.
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The Healthy Ageing and Prevention Index is an online tool created by ILC that ranks countries on six metrics including, life span, health span, work span, income, environmental performance, and happiness. The Index helps us understand how well countries have adapted to longevity and inform decision makers on what must be done to maximise the economic benefits that comes with living well for longer.
Alongside the 77th World Health Assembly in Geneva on 28 May 2024, we launched the second version of our Index, allowing us to track progress and give new insights into what needs to be done to keep populations healthier for longer.
The speakers included:
Professor Orazio Schillaci, Minister of Health, Italy
Dr Hans Groth, Chairman of the Board, World Demographic & Ageing Forum
Professor Ilona Kickbusch, Founder and Chair, Global Health Centre, Geneva Graduate Institute and co-chair, World Health Summit Council
Dr Natasha Azzopardi Muscat, Director, Country Health Policies and Systems Division, World Health Organisation EURO
Dr Marta Lomazzi, Executive Manager, World Federation of Public Health Associations
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This content provides an overview of preventive pediatrics. It defines preventive pediatrics as preventing disease and promoting children's physical, mental, and social well-being to achieve positive health. It discusses antenatal, postnatal, and social preventive pediatrics. It also covers various child health programs like immunization, breastfeeding, ICDS, and the roles of organizations like WHO, UNICEF, and nurses in preventive pediatrics.
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In this session, we will explore how a robust quality management solution can empower your organization to meet regulatory requirements and improve processes for MIPS reporting and internal quality programs. Learn how our MeasureAble application enables compliance and fosters continuous improvement.
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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/
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A Turn-Key Flow-Through-Mode Purification Process to improve Quality and Safety of Plasma IgG
1. The life science business of Merck KGaA,
Darmstadt, Germany operates as
MilliporeSigma in the U.S. and Canada.
A Turn-key flow-through-mode
purification process to improve
quality and safety of plasma IgG
Collaboration work with Taipei Medical University
Prof. Thierry Burnouf, Vice Dean, Director of the International PhD
Program in Biomedical Engineering,
College of Biomedical Engineering,
Taipei Medical University
Josephine Cheng
Senior Consultant, Core modalities APAC,
Bioprocessing strategy
Webinar Oct. 12, 2021
2. The life science business
of Merck KGaA, Darmstadt,
Germany operates as
MilliporeSigma in the U.S.
and Canada
4. Population
growth
Immunoglobulin G is the leading plasma-derived medicine
54%
16%
11%
9%
9%
IgG
Factors
Albumin
Protease Inhibitors
Others
2020 Global
Plasma Market
(28 B USD)
58%
15%
9%
10%
8%
IgG
Albumin
Factors
Protease Inhibitors
Others
2025 Global
Plasma Market
(39 B USD)
7.9%
5.4%
3.1%
7.6%
4.3%
Source: Plasma fractionation markets report 2020, marketsandmarkets.
ON/Off
label use
SCIG
Adoption
Webinar Turnkey flowthrough IGG purification | Oct. 2021
◼ Example IgG Usages:
- Primary
immunodeficiency
- Secondary
immunodeficiency
- Autoimmune and
inflammatory diseases
- Hyperimmune diseases
- Convalescent IgG (COVID)
◼ Listed as Essential
medicines by WHO
4
5. Shortage
Improvement
on Quality &
Productivity
A generic, easy-to-operate,
flowthrough-mode purification
process that provides scalable &
robust purification with enhanced
productivity and quality IGG fitting
for therapeutic usage.
High market demands drives exploration in process optimization
Webinar Turnkey flowthrough IGG purification | Oct. 2021
Accessibility
to LMIC Quality criteria:
• Virus safety
• Low IgA & IgM contamination
• Low FXI/XIa
• Lack of Hemolytic effect
• Lack of chemicals used for virus
inactivation
5
7. From Plasma donation to patient adminstration
Experimental flow with key steps used in purification
A generic, easy-to-operate,
flowthrough-mode purification
process that provides scalable
& robust purification with
enhanced productivity and
quality IGG fitting for
therapeutic usage.
Webinar Turnkey flowthrough IGG purification | Oct. 2021
Source: JC et. al. Process steps for fractionation of IgG depleted of IgA,
isoagglutinins, and devoid of in vitro thrombogenicity. Blood Transfusion 2021,
DOI 10.2450/2021.0159-21
7
8. Materials and Methods used in IgG purification process
8
No. Process step Description
1 Caprylic Acid Treatment/centrifugation
Added 5% caprylic acid to precipitate non-IgG protein and centrifuged to remove
precipitates to generate starting materials representing worse case of Fraction
I+II+III in plasma fractionation process.
2 Batch TFF UF/DF
Concentrated and diafiltrated against the chromatographic equilibration buffer
using Pellicon® 3 Biomax (30 kDa, A screen) via tangential flow filtration
(TFF) method prior to the subsequent chromatography purification steps.
3 Anion exchange (AEX) chromatography
Fractogel® EMD TMAE (M) anion exchange chromatography resin for primarily
purification. Major IgA and IgM was removed.
4
Pre-affinity chromatography SPTFF
concentration
Concentration was performed using Pellicon® 3 Biomax (30 kDa, A screen)
via Single-Pass TFF technology, to achieve an optimal loading concentration of
>40 mg/mL for following affinity chromatography purification.
5 Affinity chromatography
Eshmuno® P anti-A and anti-B, two distinct, affinity-based chromatography
resins, were used to remove blood type anti-A and anti-B isoagglutinin.
6
S/D treatment and S/D removal by C18
reverse phase chromatography
Solvent/detergent (S/D) treatment was applied to the IgG batches for enveloped
virus inactivation. The LiChroprep® RP-18 (40-63 µm) column was used in a
flow-through mode to remove the S/D.
7 SPTFF final concentration
Single-Pass TFF technology using Pellicon® 3 Biomax (30 kDa, D screen) to
achieve final target concentration of 200 mg/mL.
Webinar Turnkey flowthrough IGG purification | Oct. 2021
9. Analytical Steps involved along the process
Webinar Turnkey flowthrough IGG purification | Oct. 2021
Source: JC et. al. Process steps for fractionation of IgG depleted of IgA,
isoagglutinins, and devoid of in vitro thrombogenicity. Blood Transfusion 2021,
DOI 10.2450/2021.0159-21
9
12. Evaluating optimal chromatographic parameters for IgA and IgM
removal
Fractogel™ TMAE resin - Anion Exchange chromatography
Evaluation of pH effect on Fractogel® TMAE(M) resin binding capacity.
Typical chromatograph for a
flow-through mode anion
exchange chroamtography.
25mM Sodium Acetate buffer,
with 185cm/h flow rate.
Learn More with our webinar:
Chromatography: Chromatographic
strategies for IVIG purification - Part 2
• No detectable impact IgA/IgM binding
and IgG purification from 3 pH values.
• Optimal pH for loading identified @ pH
6.0
Webinar Turnkey flowthrough IGG purification | Oct. 2021
12
13. IgG, IgA and IgM in feed, flow-through and wash:
10 cycles (data shown previously) & 200 cycles with good
reproducibility
Webinar Turnkey flowthrough IGG purification | Oct. 2021
1. Purity IgG
increases
from 87% to
99% at small
and pilot scale.
2. Recovery avg.
96.89±7.06%
3. 200 cycles
test with
standard
cleaning
conditions
confirms the
robustness
Lab-scale anion exchange chromatography step: robustness test
13
14. IGG Subclasses distribution in 200 cycle tested to be
similar
Webinar Turnkey flowthrough IGG purification | Oct. 2021
lab-scale
Subclasses IgG 1 IgG 2 IgG3 IgG4
Feed 61.43% 34.49% 1.46% 2.62%
Flowthrough 61.83±3.35% 35.35±3.32% 1.42±0.09% 1.39±0.08%
• Satisfactory preservation of
the IgG subclass distribution.
14
15. Webinar Turnkey flowthrough IGG purification | Oct. 2021
Summary AEX:
1. Optimal pH for loading identified @ pH 6.0
2. Purity increase for IgG from 82% to 99, in
pilot scale almost 100% purity was reached.
3. 200 cycles test with standard cleaning
conditions confirms the robustness.
4. No major changes in IgG subclasses.
5. No in vitro thrombin generation detected.
Pilot scale 10 L batches study
confirms Fractogel™ as major step
removing IgA and IgM
15
17. Blood type isoaglutinins increases risk for hemolysis and
provides some limitation on plasma collection
1. High dose treatment for
Immunomodulation patients.
2. Additional safety measure for
fractionators to avoid failed batches/
decrease isoaglutinins content.
3. Safe product even for high-dose
administration.
Webinar Turnkey flowthrough IGG purification | Oct. 2021
17
18. Eshmuno® P Anti-A (FT) Eshmuno ® P Anti-B (FT)
Robust reduction of the blood-type specific isoagglutinins
Affinity chromatography
8 to 16 times reduction in Anti-A titer 16 to 32 times reduction in Anti-B titer
*samples tested at 30mg/ml concentration from 1st SPTFF (6X) step to the last step.
Webinar Turnkey flowthrough IGG purification | Oct. 2021
Feed loaded onto column with
25mM Sodium Acetate buffer
in flowthrough mode, 3 min
residence time.
18
21. Classical Solvent detergent proven to effectively reduced model
viruses with single-use bags in time as short as 5 mintues
Human IgG: 0.3% TnBP + 1% TX100 LRV Results
Virus Device
LRV at Incubation Time (min)
5 30 60 360
XMuLV
Mobius® 1 ≥ 5.5 ≥ 5.3 ≥ 5.3 ≥ 5.4
Mobius® 2 ≥ 5.5 ≥ 5.3 ≥ 5.3 ≥ 5.5
BVDV
Mobius® 1
Mobius® 2
≥ 4.5
≥ 4.4
≥ 4.4
≥ 4.6
≥ 4.6
≥ 4.4
≥ 4.5
≥ 4.5
Publication: Hsieh YT, Mullin L, Greenhalgh P, Cunningham
M, Goodrich E, et al. Single-use technology for
solvent/detergent virus inactivation of industrial plasma
products. TRANSFUSION 2016;56;1384–1393.
0.3% TnBP + 1% Triton X-100 provides > 4-5
LRV in time as short as 5 minutes
Learn More with our webinar: Solvent Detergent Viral Inactivation using
S.U Technology in Blood Fractionation Processes
Webinar Turnkey flowthrough IGG purification | Oct. 2021
21
22. Cell viability assay of 3T3 cells provides quick understanding
for S/D residual level and safety of the purified IgG
Webinar Turnkey flowthrough IGG purification | Oct. 2021
Cell toxicity data of the SD-treated IgG following C18 chromatography
Cells were treated with 0-130 ppm S/D-spiked IgG for establishment of a standard curve (A) and flow
through of S/D treated-IgG after C-18 column (B). Cells were incubated for 24 hours with the S/D-spiked
IgG or the C18 flow-through of the SD-treated IgG. The cell viability was analyzed by CCK-8 assay. FT, flow-
through; ppm, part per million; S/D, solvent/detergent.
NIH-3T3 cells treated with 2
ppm S/D-spiked IgG
(standard curve) started to
evidence a 4.3% decrease in
viability. The flow-through of
C18 (FT18) corresponding to
a loading with 18 mL per ml
of C18 packing evidenced
signs of cytotoxicity
indicating that 17 mL of S/D
spiked IgG was the
maximum volume of SD-
treated IgG to loaded onto 1
mL C18.
17ml of S/D spiked IgG/
ml packed C18 resin
When S/D concentration
~ 4ppm, viability of
cells starts to decrease
Source: JC et. al. Process steps for fractionation of IgG depleted of IgA,
isoagglutinins, and devoid of in vitro thrombogenicity. Blood Transfusion 2021,
DOI 10.2450/2021.0159-21
22
23. Efficient removal of S/D by Licroprep® C18 (40 – 63um)
sorbent to non-detectable level
Virus Inactivation and removal by C18 RP-chromatography
A. Typical chromatography for FT mode S/D-IGG running through C18 column.
B. TnBP residual tested on GC-MS with results < 1ppm, Triton X-100 residual tested by HPLC with results < 2ppm,
showing a robust removal with the loading quantity defined (6ml S/D-IGG/ml C18 packed resin), with capacity
tested at 17 ml S/D-IGG/ml C18 resin)
Residual Triton X-100 of SD-IgG
(Ratio of resin and loaded IgG)
Batch 3
(1mL:6mL)
C18 <2 ppm
SPTFF-5X <2 ppm
Residual TnBP of SD-IgG
(Ratio of resin and loaded IgG)
Batch 3
(1mL:6mL)
C18 <1 ppm
SPTFF-5X <1 ppm
A. B.
Webinar Turnkey flowthrough IGG purification | Oct. 2021
23
30. Ultrafiltation/diafiltration successfully removed caprylic
acid from the IgG fraction, data showed consistent
removal from 3 pilot scale batches
• 6x diafiltration was carried out under
constant volume mode at ~2x VCF.
• pH and conductivity from permeate were
taken at each diafiltration volume (DN). As
shown in the diafiltration profiles, target
pH and conductivity were reached at 5-
6 DN. Typically 2 DN will be added as
safety factor.
• Initial concentration : 8.74 mg/mL
• Final concentration: 16.16 mg/mL
Webinar Turnkey flowthrough IGG purification | Oct. 2021
30
31. 31
Filter
Pellicon® 3, Biomax®
30KD, A screen
Area 0.33
Initial conc. 11.76 mg/ml Initial vol. ~4 L
Final conc. 74.62 mg/ml Final vol. ~670 mL
Feed flux 0.57 LMM TMP 11-12psi
Conversion(%) ~83%
SPTFF successful concentrated the sample to 74 mg/ml, achieved optimal loading concentration
for subsequent anion exchange step. Also reduce time and volume for chromatography step.
Retentate pressure excursion Feed Flux excursion Permeate Flux TMP excursion
Single-Pass TFF optimization provides options for inline concentration
Webinar Turnkey flowthrough IGG purification | Oct. 2021
32. Webinar Turnkey flowthrough IGG purification | Oct. 2021
A gentle method for high concentration ultrafiltration
Single-Pass Tangential Flow Filtration
Summary:
1. Sequential optimization to identify
parameters to reach target concentration
20%. (200 mg/ml)
2. SPTFF before Eshmuno® P Anti-A/B step
reduced loading time, buffer consumed,
also improved the AC performance.
3. SPTFF for final concentration successfully
provided a gentle (low shear force),
scalable, high recovery, and easy-to-
operate method for high concentration
ultrafiltration.
Process Parameters
Trial volume 726 ml Feed flux 0.01 LMM
Target VCF 5 x TMP 11-12 psi
Conversion 80 %
Retentate
Pressure
~10
Final
volume
145.2 ml
LMM: feed flow rate in L per min per total square meters of membrane area;
TMP: transmembrane pressure.
32
35. TGA & Zone Electrophoresis confirming purity increases along the
process
Overall Quality test results
Webinar Turnkey flowthrough IGG purification | Oct. 2021
Thrombin generation assay (TGA) conducted on IgG fractions over the various stages of purification,
indicating Fractogel TMAE removing pro-thrombogenic factors.
Lag Phase (min) Thrombin (nM) Time to Peak (min) Velocity Index AUC
CPP 4.75 ±0.50 52.16 ± 14.73 27.25 ±19.92 6.98 ± 6.93 954.20 ± 325.20
CA-IgG 4.00 ±0.00 27.20 ±5.92 44.50 ±0.58 0.67 ± 0.15 291.24 ± 155.68
Fractogel® BDL BDL BDL BDL BDL
SPTFF BDL BDL BDL BDL BDL
*BDL= below detection limits
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Recovery from major steps (%)
35
36. Comparable purity to market product
Quality check benchmarking market product
Fractions Non-IgG IgG
Unit % %
5XSPTFF-B1 4.8 95.2
5XSPTFF-B2 1.1 98.9
5XSPTFF-B3 0.6 99.4
Privigen® IgG 0.3 99.7
Webinar Turnkey flowthrough IGG purification | Oct. 2021
Zone electrophoresis confirmed the high
purity of the final IgG. The purity of batch
3 (5X SPTFF) reaches almost 100%.
Albumin
IgG
Albumin
IgG-HC
IgG-LC
M: Protein
marker
1:CPP
2: CA-IgG
3: Fractogel ® FT
4: Anti-A-FT
5: Anti-B-FT
6: C18
7:SPTFF-5X
8:Privigen IgG
SDS-PAGE under non-reducing (left) and
reducing (right) conditions showing the
step wise purification process, purity was
enhanced especially after AEX step.
*5 µg of protein loaded on to 4-12% Bis-Tris SDS-PAGE
36
38. Conclusions
1. Milligard® PES 1.2/0.2 prefilters effectively
reduced the filtration area needed for sterile
filters e.g. Millipore Express® SHC or Durapore®
filters
2. Clarification using Millistak+® HC A1HC to
facilitate downstream purification.
3. Fractogel® TMAE(M) anion exchange
chromatography for efficient removal of IgA and
IgM, with well maintained IgG subclasses.
Proof of concept for a generic
process with intensified
processing:
4. Eshmuno® P anti-A and Eshmuno® P anti-B
chromatography for robust removal of anti-A
and anti-B agglutinins.
5. S/D used in virus inactivation can be efficiently
removed using Licroprep® C18 Reverse-phase
chromatography.
6. Use of SPTFF as a mild and robust approach to
concentrate IgG to a target of 20%
concentration.
7. Recovery from 92 – 100% for each step
resulting in an overall process recovery of
greater than 70% under a worst-case scenario,
with opportunities to improve further with
additional optimization.
Such flow-through process can be monitored reliably by controlling
key process parameters, ready to be scalable, and can be applied
for various IgG products such including polyvalent IgG,
hyperimmune, or convalescent immunoglobulins.
Webinar Turnkey flowthrough IGG purification | Oct. 2021
38
39. Publications
ISBT GBS Working Party Workshop presentation:
https://www.isbtweb.org/working-parties/global-blood-
safety/workshop-recordings
39
Blood Transfusion
publicaion:
http://www.bloodtra
nsfusion.it/articolosi
ng.aspx?id=001177
41. Acknowledgements
Taipei Medical University, Taiwan (TMU):
• Prof. Thierry Burnouf
• Yu-Wen Wu
• Chen-Yun Wang
• Cheum Lam Hong
Merck:
• Sharon ShangJung Wu
• Karen Waiyu Chan
• Leo Xun Liao
• Xisheng Cao
• Bin Wang
42. Acknowledgement to team
working behind the scene
Lynn Neild, Ravin Gami, Jessica Torres, Patryk Kelley
(MilliporeSigma, USA), Jennifer Kercher, Nina Weis,
Andreas Stein, Andre Kiesewetter, Michael Schulte
(Merck KGaA, Germany), Manuel Brantner (Merck
Chemicals and Life Science GesmbH, Austria), Anissa
Boumlic, Carole Inglevert (Millipore SAS, France),
Sandra Hon (Merck Pte Ltd, Singapore) for their support.
We thank the Taipei Blood Center for plasma supply
(Guandu, Taiwan Blood Service Foundation, Taiwan).