1) The co-op student worked at Bristol-Myers Squibb to optimize protein purification chromatography conditions and improve impurity clearance.
2) High-throughput screening was used to evaluate different buffer and resin combinations. Process conditions were optimized to separate variants and improve purity.
3) The student met project goals including developing chromatography methods, characterizing protein populations, and providing insights into improving processes. The work focused on optimizing the initial protein A capture step to reduce downstream purification needs.
Significant advances in analytical technology over the past few years have improved the quantification and characterization capabilities for subvisible ( 1 - 100 μm) and submicron particles (≤1 μm). As the technology continues to improve so do the expectations of regulatory agencies for sponsors to characterize particles in these size ranges. However, multiple orthogonal methods are required to span the entire range and accurately characterize the particle profile. Each instrument has its own limitations based on detection method and properties of therapeutic protein products that must be well understood to generate high-quality data. Written by Amber Fradkin, Ph.D. Associate Director, R&D, KBI Biopharma
Presentation at BPI West by Abhinav A. Shukla, Ph.D. Senior Vice President Development & Manufacturing KBI Biopharma, Durham NC, February 27 – March 2, 2017, Platforms for mAb Commercialization
Getting Biopharmaceutical Production Processes Right the First TimeKBI Biopharma
Strategies for rapid acceleration of cell line, upstream and downstream process development. A presentation by Ying Huang, Ph.D., Associate Director of Cell Line Development at KBI Biopharma. Presented at World Orphan Drug Congress. Washington DC. (2014)
Monoclonal antibody (mAb) therapeutics have formed and continue to form the vast majority of biopharmaceutical company pipelines today with a number of remarkable commercial successes. The advent of mAbs as therapeutics has been greatly aided by a process platform approach that has enabled rapid development and manufacturing for this class of drugs.Downstream process platforms for mAbs first evolved over a decade ago and have had a significant impact on the time and resources spent in process development. This chapter describes some of the platform approaches first used in the biopharmaceutical industry and how those platforms have evolved over the last decade based on needs as well as newly available technology. We also describe the advent of next generation mAb based constructs and the creation of possible platforms for those moieties.
Next Generation Recombinant Protein ManufacturingKBI Biopharma
Next Generation Processes: What Model Works Best to Manufacture Recombinant Proteins in Asia?
BioPharma Asia 2017
Suntec Convention Center. Singapore, March 22, 2017
Thomas Jung, M.S. Vice President, Business Development
KBI Biopharma Inc.
Significant advances in analytical technology over the past few years have improved the quantification and characterization capabilities for subvisible ( 1 - 100 μm) and submicron particles (≤1 μm). As the technology continues to improve so do the expectations of regulatory agencies for sponsors to characterize particles in these size ranges. However, multiple orthogonal methods are required to span the entire range and accurately characterize the particle profile. Each instrument has its own limitations based on detection method and properties of therapeutic protein products that must be well understood to generate high-quality data. Written by Amber Fradkin, Ph.D. Associate Director, R&D, KBI Biopharma
Presentation at BPI West by Abhinav A. Shukla, Ph.D. Senior Vice President Development & Manufacturing KBI Biopharma, Durham NC, February 27 – March 2, 2017, Platforms for mAb Commercialization
Getting Biopharmaceutical Production Processes Right the First TimeKBI Biopharma
Strategies for rapid acceleration of cell line, upstream and downstream process development. A presentation by Ying Huang, Ph.D., Associate Director of Cell Line Development at KBI Biopharma. Presented at World Orphan Drug Congress. Washington DC. (2014)
Monoclonal antibody (mAb) therapeutics have formed and continue to form the vast majority of biopharmaceutical company pipelines today with a number of remarkable commercial successes. The advent of mAbs as therapeutics has been greatly aided by a process platform approach that has enabled rapid development and manufacturing for this class of drugs.Downstream process platforms for mAbs first evolved over a decade ago and have had a significant impact on the time and resources spent in process development. This chapter describes some of the platform approaches first used in the biopharmaceutical industry and how those platforms have evolved over the last decade based on needs as well as newly available technology. We also describe the advent of next generation mAb based constructs and the creation of possible platforms for those moieties.
Next Generation Recombinant Protein ManufacturingKBI Biopharma
Next Generation Processes: What Model Works Best to Manufacture Recombinant Proteins in Asia?
BioPharma Asia 2017
Suntec Convention Center. Singapore, March 22, 2017
Thomas Jung, M.S. Vice President, Business Development
KBI Biopharma Inc.
Octet Potency Assay: Development, Qualification and Validation StrategiesKBI Biopharma
Octet Potency Assay: Development, Qualification and
Validation Strategies
Carson Cameron, Brendan Peacor, Nathan Oien, Andrew Cheeseman, and Jimmy Smedley, KBI Biopharma, Durham, NC
John Laughlin, and David O. Apiyo, ForteBio, Fremont, CA
Biopharmaceutical Formulation Development CM3 Implementation and Initial TestingKBI Biopharma
The document describes the Freeslate CM3 biologics formulation development workstation. The CM3 allows for automated sample preparation, processing, analysis, and experimental design for preformulation studies. It integrates liquid handling, analytical instruments, and software for designing and executing formulation experiments. Validation studies are performed to ensure reliable data from the liquid handling and analytical instruments. Buffer preparation and pH verification are provided as an example application.
Speed to GMP: Moving from Rapid Process Development to High Throughput Tech T...KBI Biopharma
This document discusses strategies for rapidly transferring biologics manufacturing processes from development to commercial production. It provides examples of how KBI Biopharma employs standardized platform processes and integrated development approaches to minimize changes between scales. For antibody processes, extensive use of platform cell lines, media, and unit operations allows seamless transfer. Non-antibody processes require more customization but subsequent products can still leverage a base platform. Tech transfer timelines are established early and deliverables like batch records are reviewed. This enables timely preparation for cGMP manufacturing and regulatory filings.
Application and Adaptation of Platform and Alternative Purification Steps to ...KBI Biopharma
This document discusses challenges in applying standard platform purification processes to Fc-fusion proteins. Fc-fusion proteins show more diversity than monoclonal antibodies, which can complicate rapid process development. Residue levels and yields after protein A capture can vary significantly depending on cell line and molecule properties. Alternative viral inactivation or protein A wash steps may be required. Anion exchange chromatography may require bind-and-elute mode rather than flow-through for some molecules. Ultrafiltration/diafiltration can cause unexpected pH offsets at low protein concentrations for some Fc-fusions due to Donnan and volume exclusion effects. Overcoming these challenges may require modified purification steps or formulation conditions.
HIV Vaccines Process Development & Manufacturing - Pitfalls & PossibilitiesKBI Biopharma
Originally presented at the HIV Vaccine Manufacturing Workshop –July 19th& 20th, 2017 by Abhinav A. Shukla, Ph.D.Senior Vice PresidentDevelopment & ManufacturingKBI Biopharma, Durham NC
Covering a Spectrum of New Product Development Technologies - From Cells to C...KBI Biopharma
KBI Biopharma provides analytical services, process development, and manufacturing for biologics from early discovery through commercialization. It has locations in Durham, Boulder, RTP, and The Woodlands focused on mammalian and microbial development as well as cell therapy. KBI leverages platform approaches for monoclonal antibodies, bispecific antibodies, fusion proteins, and emerging modalities. It has also partnered with Selexis to utilize their cell lines for development programs and has integrated Selexis cell lines into its platforms to rapidly develop cell lines from DNA to drug substance within 9 months.
High-throughput Miniaturized Bioreactors for Cell Culture Process Developmen...KBI Biopharma
Decreasing the timeframe for cell culture process development has been a key goal towards accelerating biopharmaceutical development. Automated Micro-scale Bioreactors (ambrTM) is an advanced micro bioreactor system with miniature single-use bioreactors with a 9-15mL working volume controlled by an automated workstation. This system was compared to conventional bioreactor systems in terms of its performance for the production of a monoclonal antibody and a non-antibody molecule in recombinant Chinese Hamster Ovary (CHO) cell lines.
The miniaturized bioreactor system was found to produce cell culture profiles that matched across scales to 3L, 15L and 200L stirred tank bioreactors. Moreover, changes to important process parameters in ambrTM resulted in predictable cell growth, viability and titer changes, which were in good agreement to historical data from the larger scales. ambrTM was found to successfully reproduce variations in temperature, dissolved oxygen and pH conditions similar to the larger bioreactor systems. Additionally, the miniature bioreactors were found to react well to perturbations in pH and dissolved oxygen through adjustments to the PID control loop. Overall, the studies demonstrate the utility of the ambrTM system as a high throughput system for cell culture process development.
High Throughput Bioreactor Mimetic in Early and Late Stage Process DevelopmentKBI Biopharma
A presentation by KBI Scientist Shahid Rameez, Ph.D. at the American Chemical Society Annual Meeting– Biochemical Technology (BIOT) Division, New Orleans, LA
Custom Affinity Chromatography for Vaccine Purification: A New PD ParadigmMilliporeSigma
Purification can account for majority of the manufacturing costs of most biological drugs. In the vaccine industry, purification processes are particularly complex with non-templated processes typically with low yields leading to higher than desired product costs.
We are actively working with industry partners to develop purification processing platform to address these challenges using affinity chromatography technologies. Such a purification platform could be amenable to diverse heterogeneous types of vaccines such as glycoconjugates, virus like particles and viruses.
In this webinar, you will learn:
-Custom affinity ligand discovery, characterization, and selection strategies
-Affinity ligand - base matrix immobilization strategies
-Performance evaluation techniques
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.
Oral presentation at ESACT 2015 (Barcelona) - Identification of process param...Albert Paul
Monoclonal antibodies (mAbs) are successful biotherapeutics in the treatment of various diseases. During manufacturing of mAbs higher molecular weight (HMW) aggregates can be formed during upstream (USP) and downstream (DSP) processing, which negatively influence product yields, reduce the therapeutic efficacy of the mAbs and trigger immunogenic responses upon administration. Reducing the level of aggregates during USP could improve the production of biopharmaceuticals and reduce the burden on expensive DSP removal of the HMW species. However, the lack of analytical tools to detect mAb aggregates in USP restricts understanding the origin of the aggregates and identifying cell culture conditions influencing product quality to reduce the level of mAb aggregates. We present a high-throughput compatible method which allows quantification of mAb aggregate formation directly in cell culture samples of Chinese hamster ovary (CHO) cells replacing falsifying, laborious and time-consuming chromatographic methods. Using this new methodology, we have screened for different culture conditions effecting mAb aggregate formation in a non-producing and a mAb producing CHO cell line. Finally, we have identified important process parameters to influencing protein aggregation in mammalian cell culture. Hence, our work demonstrates that the formation of mAb aggregates can be assessed directly in mammalian cell culture and product quality can be controlled by the selection of certain cell culture process parameters.
The Butterfly Effect: How to see the impact of small changes to your ADCMilliporeSigma
This document summarizes key aspects of characterizing antibody drug conjugates (ADCs), including:
1) A case study examining how different polyethylene glycol (PEG) linker sizes affect ADC structure and target binding. Peptide mapping by mass spectrometry showed conjugation sites varied with linker size. Hydrogen/deuterium exchange mass spectrometry showed conjugation induced conformational changes.
2) Methods for assessing ADC mechanisms of action, including measuring internalization, cytotoxicity, and effector functions.
3) An overview of MilliporeSigma's comprehensive ADC product characterization and biosafety testing services across multiple sites.
Host Cell Protein Analysis by Mass Spectrometry | KBI BiopharmaKBI Biopharma
Host Cell Protein Analysis by Mass Spectrometry. Originally presented at the 2018 Sciex Users Meeting by Michael J Nold, Ph.D., Mass Spectrometry Core Facility at KBI Biopharma.
Webinar: Evaluating Viral Clearance for Continuous ProcessesMilliporeSigma
Participate in the interactive webinar now: http://bit.ly/ViralClearanceWebinar
Is viral clearance a hurdle to implementing continuous processing? We’ll share virus spiking alternatives that may pave the way for effectively evaluating viral clearance by chromatography steps in a continuous process.
Explore our webinar library: www.emdmillipore.com/webinars
High-throughput and Automated Process Development for Accelerated Biotherapeu...KBI Biopharma
KBI Biopharma has developed high-throughput and automated processes to accelerate biotherapeutic development. This includes establishing a high-throughput process development team utilizing automated equipment and informatics solutions. Analytical case studies demonstrate automation of a residual host cell protein ELISA using a liquid handling robot, reducing analysis time from hours to minutes per sample. A second case study outlines development of a high-throughput size exclusion chromatography method, reducing run time from 30 minutes to 6 minutes while still effectively screening for high molecular weight species. These efforts allow for real-time data generation and monitoring of process development experiments.
Development and sharing of ADME/Tox and Drug Discovery Machine learning modelsSean Ekins
This document discusses the development and sharing of machine learning models for ADME/Tox prediction and drug discovery. It notes that while ADME/Tox modeling began over 15 years ago with small datasets, modern models have much larger training data and address more properties. The opportunity to get pharmaceutical companies to use open-source tools and algorithms to build and share precompetitive models is described. Examples of published models for various properties like CYP inhibition and P-gp efflux built using open descriptors and algorithms are provided. The export of models from the Collaborative Drug Discovery platform and their use in mobile apps is also covered.
This document summarizes two case studies presented by Merck Serono on using high-throughput cell culture methods to speed up media development.
In the first case study, 376 different media blends were tested in parallel fed-batch cultures to identify a new high-performance medium. Data analysis using Excel, design of experiments, and multivariate analysis identified key formulations and components. One new medium showed a 30-60% improvement in titer for three cell lines.
The second case study showed that quality attributes of products from 400 micro-scale fed-batch cultures could be analyzed to optimize critical quality attributes. A design of experiments approach tested multiple factors to match quality targets. This enabled confirmation and scale-up of an improved
Integrated utilization of high-throughput bioreactors & high-throughput analy...KBI Biopharma
There is a strong impetus towards rapidly advancing an increasing number of novel biotherapeutics to clinical trials. However, development of cell culture processes is labor intensive and time consuming. KBI focuses on a high throughput process development (HTPD) approach using high-throughput miniaturized bioreactors and high throughput analytics that generate growth, productivity and product quality data that match those seen with classical systems. This approach enables a significant reduction in the cell culture process development timeline and costs for investigational biopharmaceuticals to reach the clinic.
The document discusses developing curriculum to support careers in biomanufacturing and the bioeconomy. It proposes hands-on workshops for teachers and students to learn biomanufacturing concepts and techniques. These include transforming bacteria, purifying proteins through various types of chromatography, and analyzing purified proteins through electrophoresis. The goal is to promote STEM education and careers in the growing biomanufacturing industry.
Octet Potency Assay: Development, Qualification and Validation StrategiesKBI Biopharma
Octet Potency Assay: Development, Qualification and
Validation Strategies
Carson Cameron, Brendan Peacor, Nathan Oien, Andrew Cheeseman, and Jimmy Smedley, KBI Biopharma, Durham, NC
John Laughlin, and David O. Apiyo, ForteBio, Fremont, CA
Biopharmaceutical Formulation Development CM3 Implementation and Initial TestingKBI Biopharma
The document describes the Freeslate CM3 biologics formulation development workstation. The CM3 allows for automated sample preparation, processing, analysis, and experimental design for preformulation studies. It integrates liquid handling, analytical instruments, and software for designing and executing formulation experiments. Validation studies are performed to ensure reliable data from the liquid handling and analytical instruments. Buffer preparation and pH verification are provided as an example application.
Speed to GMP: Moving from Rapid Process Development to High Throughput Tech T...KBI Biopharma
This document discusses strategies for rapidly transferring biologics manufacturing processes from development to commercial production. It provides examples of how KBI Biopharma employs standardized platform processes and integrated development approaches to minimize changes between scales. For antibody processes, extensive use of platform cell lines, media, and unit operations allows seamless transfer. Non-antibody processes require more customization but subsequent products can still leverage a base platform. Tech transfer timelines are established early and deliverables like batch records are reviewed. This enables timely preparation for cGMP manufacturing and regulatory filings.
Application and Adaptation of Platform and Alternative Purification Steps to ...KBI Biopharma
This document discusses challenges in applying standard platform purification processes to Fc-fusion proteins. Fc-fusion proteins show more diversity than monoclonal antibodies, which can complicate rapid process development. Residue levels and yields after protein A capture can vary significantly depending on cell line and molecule properties. Alternative viral inactivation or protein A wash steps may be required. Anion exchange chromatography may require bind-and-elute mode rather than flow-through for some molecules. Ultrafiltration/diafiltration can cause unexpected pH offsets at low protein concentrations for some Fc-fusions due to Donnan and volume exclusion effects. Overcoming these challenges may require modified purification steps or formulation conditions.
HIV Vaccines Process Development & Manufacturing - Pitfalls & PossibilitiesKBI Biopharma
Originally presented at the HIV Vaccine Manufacturing Workshop –July 19th& 20th, 2017 by Abhinav A. Shukla, Ph.D.Senior Vice PresidentDevelopment & ManufacturingKBI Biopharma, Durham NC
Covering a Spectrum of New Product Development Technologies - From Cells to C...KBI Biopharma
KBI Biopharma provides analytical services, process development, and manufacturing for biologics from early discovery through commercialization. It has locations in Durham, Boulder, RTP, and The Woodlands focused on mammalian and microbial development as well as cell therapy. KBI leverages platform approaches for monoclonal antibodies, bispecific antibodies, fusion proteins, and emerging modalities. It has also partnered with Selexis to utilize their cell lines for development programs and has integrated Selexis cell lines into its platforms to rapidly develop cell lines from DNA to drug substance within 9 months.
High-throughput Miniaturized Bioreactors for Cell Culture Process Developmen...KBI Biopharma
Decreasing the timeframe for cell culture process development has been a key goal towards accelerating biopharmaceutical development. Automated Micro-scale Bioreactors (ambrTM) is an advanced micro bioreactor system with miniature single-use bioreactors with a 9-15mL working volume controlled by an automated workstation. This system was compared to conventional bioreactor systems in terms of its performance for the production of a monoclonal antibody and a non-antibody molecule in recombinant Chinese Hamster Ovary (CHO) cell lines.
The miniaturized bioreactor system was found to produce cell culture profiles that matched across scales to 3L, 15L and 200L stirred tank bioreactors. Moreover, changes to important process parameters in ambrTM resulted in predictable cell growth, viability and titer changes, which were in good agreement to historical data from the larger scales. ambrTM was found to successfully reproduce variations in temperature, dissolved oxygen and pH conditions similar to the larger bioreactor systems. Additionally, the miniature bioreactors were found to react well to perturbations in pH and dissolved oxygen through adjustments to the PID control loop. Overall, the studies demonstrate the utility of the ambrTM system as a high throughput system for cell culture process development.
High Throughput Bioreactor Mimetic in Early and Late Stage Process DevelopmentKBI Biopharma
A presentation by KBI Scientist Shahid Rameez, Ph.D. at the American Chemical Society Annual Meeting– Biochemical Technology (BIOT) Division, New Orleans, LA
Custom Affinity Chromatography for Vaccine Purification: A New PD ParadigmMilliporeSigma
Purification can account for majority of the manufacturing costs of most biological drugs. In the vaccine industry, purification processes are particularly complex with non-templated processes typically with low yields leading to higher than desired product costs.
We are actively working with industry partners to develop purification processing platform to address these challenges using affinity chromatography technologies. Such a purification platform could be amenable to diverse heterogeneous types of vaccines such as glycoconjugates, virus like particles and viruses.
In this webinar, you will learn:
-Custom affinity ligand discovery, characterization, and selection strategies
-Affinity ligand - base matrix immobilization strategies
-Performance evaluation techniques
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.
Oral presentation at ESACT 2015 (Barcelona) - Identification of process param...Albert Paul
Monoclonal antibodies (mAbs) are successful biotherapeutics in the treatment of various diseases. During manufacturing of mAbs higher molecular weight (HMW) aggregates can be formed during upstream (USP) and downstream (DSP) processing, which negatively influence product yields, reduce the therapeutic efficacy of the mAbs and trigger immunogenic responses upon administration. Reducing the level of aggregates during USP could improve the production of biopharmaceuticals and reduce the burden on expensive DSP removal of the HMW species. However, the lack of analytical tools to detect mAb aggregates in USP restricts understanding the origin of the aggregates and identifying cell culture conditions influencing product quality to reduce the level of mAb aggregates. We present a high-throughput compatible method which allows quantification of mAb aggregate formation directly in cell culture samples of Chinese hamster ovary (CHO) cells replacing falsifying, laborious and time-consuming chromatographic methods. Using this new methodology, we have screened for different culture conditions effecting mAb aggregate formation in a non-producing and a mAb producing CHO cell line. Finally, we have identified important process parameters to influencing protein aggregation in mammalian cell culture. Hence, our work demonstrates that the formation of mAb aggregates can be assessed directly in mammalian cell culture and product quality can be controlled by the selection of certain cell culture process parameters.
The Butterfly Effect: How to see the impact of small changes to your ADCMilliporeSigma
This document summarizes key aspects of characterizing antibody drug conjugates (ADCs), including:
1) A case study examining how different polyethylene glycol (PEG) linker sizes affect ADC structure and target binding. Peptide mapping by mass spectrometry showed conjugation sites varied with linker size. Hydrogen/deuterium exchange mass spectrometry showed conjugation induced conformational changes.
2) Methods for assessing ADC mechanisms of action, including measuring internalization, cytotoxicity, and effector functions.
3) An overview of MilliporeSigma's comprehensive ADC product characterization and biosafety testing services across multiple sites.
Host Cell Protein Analysis by Mass Spectrometry | KBI BiopharmaKBI Biopharma
Host Cell Protein Analysis by Mass Spectrometry. Originally presented at the 2018 Sciex Users Meeting by Michael J Nold, Ph.D., Mass Spectrometry Core Facility at KBI Biopharma.
Webinar: Evaluating Viral Clearance for Continuous ProcessesMilliporeSigma
Participate in the interactive webinar now: http://bit.ly/ViralClearanceWebinar
Is viral clearance a hurdle to implementing continuous processing? We’ll share virus spiking alternatives that may pave the way for effectively evaluating viral clearance by chromatography steps in a continuous process.
Explore our webinar library: www.emdmillipore.com/webinars
High-throughput and Automated Process Development for Accelerated Biotherapeu...KBI Biopharma
KBI Biopharma has developed high-throughput and automated processes to accelerate biotherapeutic development. This includes establishing a high-throughput process development team utilizing automated equipment and informatics solutions. Analytical case studies demonstrate automation of a residual host cell protein ELISA using a liquid handling robot, reducing analysis time from hours to minutes per sample. A second case study outlines development of a high-throughput size exclusion chromatography method, reducing run time from 30 minutes to 6 minutes while still effectively screening for high molecular weight species. These efforts allow for real-time data generation and monitoring of process development experiments.
Development and sharing of ADME/Tox and Drug Discovery Machine learning modelsSean Ekins
This document discusses the development and sharing of machine learning models for ADME/Tox prediction and drug discovery. It notes that while ADME/Tox modeling began over 15 years ago with small datasets, modern models have much larger training data and address more properties. The opportunity to get pharmaceutical companies to use open-source tools and algorithms to build and share precompetitive models is described. Examples of published models for various properties like CYP inhibition and P-gp efflux built using open descriptors and algorithms are provided. The export of models from the Collaborative Drug Discovery platform and their use in mobile apps is also covered.
This document summarizes two case studies presented by Merck Serono on using high-throughput cell culture methods to speed up media development.
In the first case study, 376 different media blends were tested in parallel fed-batch cultures to identify a new high-performance medium. Data analysis using Excel, design of experiments, and multivariate analysis identified key formulations and components. One new medium showed a 30-60% improvement in titer for three cell lines.
The second case study showed that quality attributes of products from 400 micro-scale fed-batch cultures could be analyzed to optimize critical quality attributes. A design of experiments approach tested multiple factors to match quality targets. This enabled confirmation and scale-up of an improved
Integrated utilization of high-throughput bioreactors & high-throughput analy...KBI Biopharma
There is a strong impetus towards rapidly advancing an increasing number of novel biotherapeutics to clinical trials. However, development of cell culture processes is labor intensive and time consuming. KBI focuses on a high throughput process development (HTPD) approach using high-throughput miniaturized bioreactors and high throughput analytics that generate growth, productivity and product quality data that match those seen with classical systems. This approach enables a significant reduction in the cell culture process development timeline and costs for investigational biopharmaceuticals to reach the clinic.
The document discusses developing curriculum to support careers in biomanufacturing and the bioeconomy. It proposes hands-on workshops for teachers and students to learn biomanufacturing concepts and techniques. These include transforming bacteria, purifying proteins through various types of chromatography, and analyzing purified proteins through electrophoresis. The goal is to promote STEM education and careers in the growing biomanufacturing industry.
Learn about the latest innovations at MilliporeSigmaMilliporeSigma
The document advertises and provides details about technical presentations and posters being presented by MilliporeSigma and their partners at the 253rd ACS Annual Meeting from April 2-6, 2017 in San Francisco, CA. It lists over a dozen presentations covering topics like tangential flow filtration, flocculation, chromatography, high viscosity processing, continuous processing, and viral filtration. Details are provided for each presentation including date, time, location, presenters, and abstract. The poster also provides contact information for MilliporeSigma.
- Assay development is the process of creating biological and compound screening assays to identify compounds, called "hits", that have desired activity at drug targets. This involves developing biochemical and cell-based assays.
- Key factors in assay development include relevance, reproducibility, quality as measured by Z'-factor, and avoiding interference. High throughput screening uses automation to test tens of thousands of compounds against targets daily using miniaturized assays.
- Biochemical assays use purified protein or enzyme targets, while cell-based assays examine responses at transcriptional, proliferation, or second messenger levels. Automation and robotics are important for achieving desired screening rates in high throughput screening.
Savannah Neu_Senior Design Problem StatementSavannah Neu
Immucor, a biotechnology company, is experiencing decreases in the quality and quantity of HLA protein produced from their B cell culture process. They have asked the A-Team to identify the root cause of this production problem and propose solutions to increase HLA and B cell yield. The A-Team envisions redesigning cell culture protocols, constructing a system to monitor cell growth in real-time, and implementing new organization systems to promote continuous improvement. Their proposed solution involves characterizing the B cell lines, analyzing current production systems, and collecting data on cell health and growth variables to quantify HLA protein production, staying within a $3,000 cost.
Ronald Preibis has over 17 years of experience in the biotech field, including extensive experience in all aspects of cell culture and purification. He has worked in manufacturing roles at several biotech companies, leading teams and supervising processes. His experience includes process development, cGMP manufacturing, analytical testing, documentation, and ensuring regulatory compliance. He is pursuing additional education in life sciences and biomedical laboratory sciences.
Patrick Flanagan has over 15 years of experience in formulation development, analytical chemistry, and quality control. He currently works as a Principal Research Associate at Genzyme, a Sanofi company, where he supervises multiple employees and has hands-on experience in cGMP operations. Flanagan has led cross-functional teams, coordinated meetings and resources, and represented his department at internal and external conferences. He has extensive experience characterizing and developing formulations for various protein therapeutics.
Rilas Technologies has developed a universal purification process for compound libraries that removes purification as a bottleneck. The process uses exploratory analytical methods to guide selection of an appropriate preparative HPLC method. Samples are analyzed in 6 minutes and a focused prep gradient is selected in 2 minutes. The preparative run takes 10 minutes and fractions are analyzed in 6 minutes each. Within a day, a 24 compound library can be purified and ready for biological testing. In case studies, purification success rates of over 75% were achieved for random compound libraries purified without any structural information. A 240 compound library was purified within 3 weeks, with 204 compounds passing quality criteria.
Janet Willis has over 10 years of experience in analytical chemistry and quality assurance. She holds a B.S. in Chemistry from UC San Diego and has worked at various pharmaceutical companies performing analytical testing, method development and validation, quality control, and quality assurance activities. Her experience includes HPLC, GC, UV-Vis, ELISA testing and method troubleshooting. She is proficient in GLP, GCP, and cGMP regulations and analytical software programs.
A Review on Chromatography-based purification of monoclonal antibodyIRJET Journal
This document discusses chromatography-based purification methods for monoclonal antibodies (mAbs). It begins with an introduction to mAbs and their importance as therapeutic agents. The key steps in downstream processing and purification of mAbs are outlined. Protein A chromatography is described as the most common capture method due to its high selectivity and ability to remove contaminants. Additional chromatography techniques used for polishing include ion exchange and multimodal chromatography. The mechanisms and applications of each technique are summarized. In conclusion, chromatography remains the foundation of mAb purification due to properties like scalability, robustness and selectivity.
The document discusses ways to accelerate vaccine development and manufacturing. It proposes adopting a template and platform approach to streamline process development. A template provides a standardized starting point for each vaccine's development process. A platform accumulates expertise across multiple vaccines using common unit operations, parameters, and facilities. This approach can speed development times, lower costs, simplify supply chains, and facilitate technology transfer and manufacturing.
George Baklayan has over 25 years of experience in the pharmaceutical industry, holding positions at companies such as ISTA Pharmaceuticals, Xoma Ltd., University of Southern California, and California Institute of Technology. He has a proven track record of successfully developing and launching multiple drug products from concept through approval. His expertise includes analytical method development and validation, process development, pre-clinical studies, and regulatory filings.
Roadmap for Drug Product Development and Manufacturing of Biologics.pptxChintan Kalsariya
The development of therapeutic biologics involves a streamlined approach for their formulation and drug product development from early stages to process validation and commercialization.
This roadmap is based on experience with approved products and aims to improve safety, efficacy, and immunogenicity profiles in human patients, as well as maintain consistently high quality, efficiency, and reduced cost.
The approach should be applicable across all biotherapeutic products.
This document contains a summary of Mehul D. Patel's work experience and qualifications. It lists his current role as a Research Associate at Torrent Pharmacuticles Ltd since 2006, where he conducts bioanalytical activities and drug metabolism/pharmacokinetics research. Previously he worked as a Research Associate at CADILA Pharmaceutical Ltd from 2005-2006 developing and validating analytical methods. He has a M.Pharm in Pharmacology and B.Pharm degree, and has over 15 years of experience in bioanalytical method development and validation using LC-MS/MS and HPLC techniques.
The document discusses the process of developing, optimizing, characterizing, and commercializing a pharmaceutical product. It involves designing a manufacturing process to consistently deliver the intended effects of the drug product. Process development includes determining facility, equipment, materials, procedures, and validation. Optimization compares lead compounds to select those with the greatest potential to be safe and effective medicines. Characterization tests understand the physical and chemical properties of materials. Commercialization requires approvals from regulators and establishing manufacturing, distribution, and marketing capabilities to introduce the product into markets. The goal is to produce a drug that is safe, effective, and affordable to improve patient health.
High throughput screening is a type of assay. By this assay we can identified the target or binding site of drugs. Its mainly performed during the drug discovery process.
The document is a resume for Kanagasabapathi S., highlighting his expertise and 10+ years of experience in bioanalytical method development and drug discovery. It summarizes his responsibilities developing over 1000 analytical methods for small molecules, conducting ADME assays, and managing projects. Currently he works as a Research Scientist at Syngene International Ltd, where he leads projects and trains junior scientists.
The document discusses various stages and techniques involved in downstream processing. Downstream processing refers to the recovery and purification of biosynthetic products from natural sources like fermentation broth. It involves removing insolubles, isolating the product, purifying the product, and final polishing. Key techniques discussed include filtration, centrifugation, chromatography, cell disruption through mechanical or non-mechanical means like homogenization, and analytical support through techniques like LC-MS peptide mapping.
Purification method development for chiral separation in supercritical
fluid chromatography with the solubilities in supercritical fluid
chromatographic mobile phases
1) ESW-Biodiesel is a student project at RPI that converts used vegetable oil into biodiesel on a large scale to provide hands-on experience for chemical engineering students.
2) The process involves collecting used vegetable oil from campus dining halls, converting it into biodiesel via a transesterification reaction, and providing the biodiesel to customers in the community.
3) The project aims to develop a more effective and optimized large-scale biodiesel production process through collaboration between students and academic partners.
The report details the Engineers for a Sustainable World (ESW) - RPI Chapter's biodiesel project. The project aims to convert waste vegetable oil from campus dining halls into biodiesel that can power campus shuttles. The project leaders have developed a small-scale testing procedure and are seeking funding for a 200-gallon batch processing facility to be located in Blaw Knox. The long term goals include obtaining a vehicle that can run on the produced biodiesel in order to promote sustainability on campus and support future ESW projects.
The document discusses optimization of monoclonal antibody purification using protein-A chromatography and precipitation methods. It describes how precipitation of cell culture harvest at low pH can selectively remove impurities. It also discusses how manipulating pH and additives in protein-A chromatography wash buffers can further reduce impurity levels. Optimizing the neutralization pH after protein-A elution can also selectively remove impurities. The optimization strategy reduced process impurities like HCPs and DNA to meet drug substance specifications using just one chromatography column.
- The document describes the intern's work testing and integrating sounding rocket payloads at Orbital Sciences Corporation under NASA's Sounding Rocket Operations Contract.
- Key tasks included aiding the testing of payloads through procedures like vibration, bend, and balance testing to ensure durability and mission success.
- The intern also helped design hardware components and gained an understanding of manufacturing processes.
- Overall the internship met the goals of learning about payload integration timelines and systems, and developing mechanical engineering skills.
This document provides a final report on an internship with NASA's RESOLVE project. The report summarizes testing done on various components of the RESOLVE Engineering Test Unit (ETU) fluid subsystem. Pressure transducers were tested for repeatability under varying pressure and temperature conditions. Resistance temperature detectors were tested using a procedure that exposed them to a range of temperatures in atmospheric and vacuum environments. Additional tests analyzed heat wrapping configurations, compared options for gold deposition, and evaluated a mini-manifold design. The results of these tests will help optimize components for integrated testing of the ETU fluid subsystem and other RESOLVE subsystems in a vacuum chamber. The internship provided hands-on experience with testing spaceflight hardware and procedures
The FSS is currently undergoing engineering testing to improve design reliability for its flight mission. Testing includes pressure transducer repeatability under temperature changes, heat wrap efficiency for heating tubing, and thermal profiling of manifolds. Pressure transducer testing showed reliable performance under temperature and pressure changes. Heat wrap testing found that three wraps per inch minimizes heat loss and uneven heating. Thermal profiling found uneven heating across tubing, with ends and areas near manifolds cooler due to heat drawing away.
1. Bristol-Myers Squibb (BMS) – Rensselaer Polytechnic Institute (RPI) – Cooperative Education Work Report
Rensselaer Polytechnic Institute i 12/3/2013
Cooperative Education Project Work Report
Rensselaer Polytechnic Institute
Evaluation of Protein-A Wash Conditions for Antibody Purification
Duration: 13JAN14 – 03JUL14
Ray Parker1
Rensselaer Polytechnic Institute, Bachelor of Science, Chemical Engineering
Bristol-Myers Squibb, Hopkinton, Massachusetts 01748
Mentors: Srinivas Chollangi Ph.D.2
, Yi Li Ph.D.3
1
Downstream Process Development Co-op, Chemical Engineering, Rensselaer Polytechnic Institute.
2
Scientist I, Chemical Engineering, Bristol-Myers Squibb, University of Oklahoma – Norman, University of Texas.
3
Senior Scientist, Chemical Engineering, University of Delaware – Newark, University of Arkansas
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I. Part One
Bristol-Myers Squibb is a specialized biopharmaceutical company that focuses on the discovery,
commercialization process development, clinical trials and commercialization of new drugs. Bristol-Myers Squibb
manufactures prescription pharmaceuticals in several therapeutic areas, such as cancer, HIV/AIDS, cardiovascular
disease, diabetes, hepatitis, rheumatoid arthritis and psychiatric disorders. Bristol-Myers Squibb’s mission is to
discover, develop and deliver innovative medicines that help patients prevail over serious diseases.
The entire pipeline process starts in the Cell Line Development, where each specific cell line that codes for the
specific antibody is developed. Then the antibody must be purified and a process must be developed in order to purify
the antibody to be injected into patients. Development of the commercial purification process falls to the responsibility
of the Process Development Department. Once a purification protocol is finalized, the drug substance must undergo
testing in the Clinical Trials Phase. Once extensive studies have been completed on the antibody substance, the drug
can then move onto commercialization development to be manufactured and then sold to patients around the world.
Co-op Abstract. Under the direction of BMS mentor, the Co-Op will work with the
Downstream Process Development team to develop novel separation technology and gain further
understanding of the current protein purification platform. The project will focus on key unit operations
such as ion-exchange, hydrophobic, and mixed mode chromatography to improve impurity clearance
and product yield. Experimental work will include process condition optimization, process
characterization, as well as process understanding at the molecular level. The Co-Op will collaborate
with BMS TECAN team (from multiple sites) to implement high throughput screening tools and will
use design of experiments (DOE) methodology to map out operational space. In addition, biophysical
characterization of glycoprotein structure will be performed to assist process understanding.
A. Key Deliverables at Project Completion:
1. Develop chromatography conditions to separate charge variants, aggregate populations, and
different glycol-forms.
2. Characterize heterogeneous protein populations and impurities (when applicable) with various
biophysical and biochemistry tools.
Cell Line Development Harvest Development Purification Process Final Drug Substance
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3. Study mechanistic and empirical correlations between process parameters and product attributes
and provide insight to current process issues (such as impurity clearance and product stability).
B. Key Learning:
1. Protein bio-separation techniques in an industrial setting.
2. Process analytical and characterization tools.
3. Downstream process development essential knowledge.
Process Development or PD, focuses on the developing methods for expressing and purifying the drug substance
to move onto formulation. PD consists of two sub-departments; Process Development Upstream and Process
Development Downstream. Process Development Upstream uses the cell line developed by the Cell Line
Development Department to optimize host cell growth and antibody/titer production to produce harvest, which is a
mixture of cell debris and antibodies. The harvest is then processed in Downstream Process Development, which
removes host cell proteins and other impurities.
Downstream Process Development helps to ensure that the quality of the drug remains consistent and robust. The
process in Downstream Process Development goes from Clarification (Centrifugation/Depth Filtration), to Capture
Chromatography (Protein-A/Viral Inactivation) to Intermediate Purification Chromatography to Polishing
Chromatography then to Viral Filtration. For each of the chromatography steps, there are sanitization, equilibration
and load steps. Then following the load step, there are a various number of wash steps, which are then followed by
elution, strip/regeneration and then sanitization, neutralization and storage of the column. During the eluate phase, the
elution is collected and processed further down the pipeline.
Strategies for separating monoclonal antibody are changing from molecule specific procedures to platform
strategies. With the help of high-throughput testing strategies platform process has become increasingly robust and
effective, removing more process and product related impurities such as DNA, HCP, CHO Cells & High Molecular
Weight (HMW) and Low Molecular Weight (LMW), respectively. The Capture Step (Protein-A), or affinity
chromatography, is the first step in downstream process development, which involves binding to the target antibody.
By effectively optimizing Protein-A, we can reduce the workload on the other columns downstream of the Capture
Step, helping to make the process more robust and efficient, which can reduce the amount of chromatography steps
required, saving time and money for BMS.
In order to find out which buffer/resin/molecule combinations are most effective in removing non-specific binding
interactions between HCP’s and mAb’s, screening tools were used.
High-Throughput Screening Optimization
• For each chromatography condition, buffer conditions were made with a multi-channel pipette in a separate
96 well plate before testing. Harvest material was then filtered and prepared for Protein-A chromatography.
The PreDictor resin plates were equilibrated with equilibration and sanitization buffer. The protein, Anti-
Lag3, was then loaded onto the resin. The resin was washed with equilibration buffer, the variable excipients
wash, and another wash step to remove any unbound material. The column was eluted and collected and
then stripped with a harsher buffer. The column was cleaned with strip buffer and stored in alcohol. Tests
were repeated with different buffers that had different pH values of 5.5, 7.0 and 9.0.
Column Screening Optimization
• The Wash pH Study consists of a similar protocol as the Wash Excipients Study, but rather than preparing
different excipients washes, different wash buffers at pH values ranging from 3.5 to 8.5 (in .5 increments)
were used to see the effects of pH on each molecule.
Viral Inactivation Process Optimization
• The Viral Inactivation Study deters the optimal pH that removes the most Host Cell Proteins (HCP) and
other aggregates. Protein A elution was titrated down to pH 3.5, held for one hour, then brought back up.
Results were then analyzed for DNA, Titer and HCP.
Analytical Techniques
• Chromatograms were run on GE AKTA™ Avants. Turbidity and UV readings were tested in Agilent
Technologies Cary 60 UV-Vis at A280nm & A410nm. Monomer content of the pooled flow through fractions
was analyzed by size exclusion chromatography using Acquity UPLC. ELISA Assays were run using
TECAN Freedom EVO®
. Statistical analyses were performed using Microsoft®
Excel and SAS JMP 8.0.
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The goal for the duration of this co-op rotation was to identify Wash and Viral Inactivation strategies that improve
impurity clearance of Protein-A capture Chromatography, making the process more robust and efficient. This is
important because the Capture step is one of the most important steps since it is early on in the purification process,
and can potentially reduce the total number of chromatography steps required after Protein-A, which can save money
and time for BMS. With the reduction of impurities, it also reduces the load on the later downstream development
chromatography steps making the process more consistent and effective.
II. Part Two
In comparison to the job description and my expectations at BMS, I was pleased. I expected to be able to;
Develop chromatography conditions to separate charge variants, aggregate, and different glycoforms.
Characterize heterogeneous protein populations and impurities with various biophysical/biochemistry tools.
Study mechanistic and empirical correlations between process parameters and product attributes and provide
insight to current process issues (such as impurity clearance and product stability).
Learn about Upstream and Downstream Process
Learn at least one new computer software/application.
Be able to describe the entire process development process.
Be able to present well on material by the end of the co-op
Be a contributor to a publication in while on co-op.
All of my goals have been met, besides being a contributor to a publication while on co-op; this may happen later
on. My mentor, Srinivas Chollangi, has been very patience while working with me on the concepts of Process
Development, which do not come naturally for me. In addition to this, my colleagues that I share laboratory space
with, have been very helpful in my development as a scientist too. They have all given me suggestions on how to do
things differently to improve my skills as well as given me conceptual advice on what could be done more effectively
while running experiments and other concepts in Downstream Process Development.
The main objective of my co-op project was to reduce the amount of chromatography steps required to purify
certain drug substances in order to reduce costs, since chromatography resin very expensive (upwards to $15,000 for
one liter). During the project, I was able to optimize chromatography processes for one capture step so effectively that
we were able to get the same purity with one column and one filter that the older process was able to achieve with
three columns and one filter. When scaling up to a commercial process, this could reduce costs.
In addition to learning about Downstream Process Development, I have had the opportunity to work in other field
as well, such as in Upstream Process Development and the Process Development Analytics Testing Group as well. A
list of skills learned and concepts introduced are listed below. In addition to these skills, I have accomplished much
during my time at BMS, including; making over 400 buffers, running over 100 Avant runs, helping on five different
molecules, running over 25 DBC tests, running over ~6 studies and <60 different experiments, packing over ~50
columns, and helping over 10 colleagues with testing/buffers/sample prep.
Downstream PD Concepts
• GE Pilot, Avant, & Explorer Usage
• Column Packing & Testing
• Buffer Making and Preparation
• Protein-A Resins Usage
• AEX/CEX Resins Usage
• HIC Resins Usage
• Membrane Chromatography
• Resin Amount Calculation
• Residence Time Calculations
• High-Throughput Screening Tools
• Amount of Harvest to Load
• DOE Experiments Testing
• Viral Filtration Setup
• Viral Inactivation Process
• Tangential Flow Filtration Setup
Software
• GE UNICORN Software
• Multi-Genetic Algorithms (MGA)
ADT Concepts
• ELISA - Dilution Plates Preparation
• A410 Testing
• A280 Testing
• HPLC/UPLC Exposure
Miscellaneous Concepts
• 6S Orientation Preparation
• GMP/Laboratory Safety Training
• Sample Data Mining
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No patents or honors/rewards were received. Various publications and presentations may represent my work in
the future.
Rensselaer’s Co-op Program has been very insightful and has been a great learning experience. I am thankful for
the opportunity to be able to go on co-op. My only comment to make the co-op program better would be to have the
monthly emails sent out on time at the beginning of the month. I cannot think of anything different.
Overall, my experience at Bristol-Myers Squibb has been very interesting and useful to my future. I have never thought
that I would be able to have a clear understanding of how some pharmaceutical companies work, or the basic process
behind how certain mAb drug substances are made. In addition to gaining an understanding of the process, I’ve been
able to help accomplish very important goals that will have a fairly large impact on the company and my work will
help to reduce costs and save time for BMS when developing drug molecules that I have been working on.