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Engineered antibodies currently represent over 30% of biopharmaceuticals in clinical trials and their total worldwide sales continue to increase significantly. The importance of antibody applications is reflected in their increasing clinical and industrial applications as well as in the progression of established and emerging production strategies. This volume provides detailed coverage of the generation, optimization, characterization, production and applications of antibody. It provides the necessary theoretical background and description of methods for the expression of antibody in microbial and animal cell cultures and in transgenic animals and plants. There is a strong focus on those issues related to the production of intrabodies, bispecific antibody and antibody fragments and also to novel applications in cancer immunotherapy.
Book Synopsis Antibody Expression and Production by : Mohamed Al-Rubeai
Download or read book Antibody Expression and Production written by Mohamed Al-Rubeai and published by Springer Science & Business Media. This book was released on 2011-05-16 with total page 349 pages. Available in PDF, EPUB and Kindle. Book excerpt: Engineered antibodies currently represent over 30% of biopharmaceuticals in clinical trials and their total worldwide sales continue to increase significantly. The importance of antibody applications is reflected in their increasing clinical and industrial applications as well as in the progression of established and emerging production strategies. This volume provides detailed coverage of the generation, optimization, characterization, production and applications of antibody. It provides the necessary theoretical background and description of methods for the expression of antibody in microbial and animal cell cultures and in transgenic animals and plants. There is a strong focus on those issues related to the production of intrabodies, bispecific antibody and antibody fragments and also to novel applications in cancer immunotherapy.
The American Anti-Vivisection Society (AAVS) petitioned the National Institutes of Health (NIH) on April 23, 1997, to prohibit the use of animals in the production of mAb. On September 18, 1997, NIH declined to prohibit the use of mice in mAb production, stating that "the ascites method of mAb production is scientifically appropriate for some research projects and cannot be replaced." On March 26, 1998, AAVS submitted a second petition, stating that "NIH failed to provide valid scientific reasons for not supporting a proposed ban." The office of the NIH director asked the National Research Council to conduct a study of methods of producing mAb. In response to that request, the Research Council appointed the Committee on Methods of Producing Monoclonal Antibodies, to act on behalf of the Institute for Laboratory Animal Research of the Commission on Life Sciences, to conduct the study. The 11 expert members of the committee had extensive experience in biomedical research, laboratory animal medicine, animal welfare, pain research, and patient advocacy (Appendix B). The committee was asked to determine whether there was a scientific necessity for the mouse ascites method; if so, whether the method caused pain or distress; and, if so, what could be done to minimize the pain or distress. The committee was also asked to comment on available in vitro methods; to suggest what acceptable scientific rationale, if any, there was for using the mouse ascites method; and to identify regulatory requirements for the continued use of the mouse ascites method. The committee held an open data-gathering meeting during which its members summarized data bearing on those questions. A 1-day workshop (Appendix A) was attended by 34 participants, 14 of whom made formal presentations. A second meeting was held to finalize the report. The present report was written on the basis of information in the literature and information presented at the meeting and the workshop.
Book Synopsis Monoclonal Antibody Production by : National Research Council
Download or read book Monoclonal Antibody Production written by National Research Council and published by National Academies Press. This book was released on 1999-05-06 with total page 74 pages. Available in PDF, EPUB and Kindle. Book excerpt: The American Anti-Vivisection Society (AAVS) petitioned the National Institutes of Health (NIH) on April 23, 1997, to prohibit the use of animals in the production of mAb. On September 18, 1997, NIH declined to prohibit the use of mice in mAb production, stating that "the ascites method of mAb production is scientifically appropriate for some research projects and cannot be replaced." On March 26, 1998, AAVS submitted a second petition, stating that "NIH failed to provide valid scientific reasons for not supporting a proposed ban." The office of the NIH director asked the National Research Council to conduct a study of methods of producing mAb. In response to that request, the Research Council appointed the Committee on Methods of Producing Monoclonal Antibodies, to act on behalf of the Institute for Laboratory Animal Research of the Commission on Life Sciences, to conduct the study. The 11 expert members of the committee had extensive experience in biomedical research, laboratory animal medicine, animal welfare, pain research, and patient advocacy (Appendix B). The committee was asked to determine whether there was a scientific necessity for the mouse ascites method; if so, whether the method caused pain or distress; and, if so, what could be done to minimize the pain or distress. The committee was also asked to comment on available in vitro methods; to suggest what acceptable scientific rationale, if any, there was for using the mouse ascites method; and to identify regulatory requirements for the continued use of the mouse ascites method. The committee held an open data-gathering meeting during which its members summarized data bearing on those questions. A 1-day workshop (Appendix A) was attended by 34 participants, 14 of whom made formal presentations. A second meeting was held to finalize the report. The present report was written on the basis of information in the literature and information presented at the meeting and the workshop.
If the antibody industry is to achieve its full potential in the next decade, the individual technical potentials must be exploited, the limitations must be addressed, and lessons learned must be applied both to current purification methods and to the new technologies that continue to emerge. This book presents an overview of the current advances applied in the manufacture of monoclonal antibody including: -concepts in development of manufacturing strategies, -importance of antibody fragments, -application of chromatography method development, -quality control, -effect of expression on antibody properties, -virus removal and safety, -pharmacokinetics, -regulatory aspects.
Book Synopsis Antibodies by : G. Subramanian
Download or read book Antibodies written by G. Subramanian and published by Springer Science & Business Media. This book was released on 2013-03-07 with total page 260 pages. Available in PDF, EPUB and Kindle. Book excerpt: If the antibody industry is to achieve its full potential in the next decade, the individual technical potentials must be exploited, the limitations must be addressed, and lessons learned must be applied both to current purification methods and to the new technologies that continue to emerge. This book presents an overview of the current advances applied in the manufacture of monoclonal antibody including: -concepts in development of manufacturing strategies, -importance of antibody fragments, -application of chromatography method development, -quality control, -effect of expression on antibody properties, -virus removal and safety, -pharmacokinetics, -regulatory aspects.
Integrating advances in molecular biology into bioprocesses presents a continuous challenge to scientists and bioengineers. This series is conceived to help meet this challenge. It examines and assesses the feasibility of new approaches for the modification of cellular function such as gene expression, protein processing, secretion, glycosylation, immortalisation, proliferation, and apoptosis as well as the systematic study of the metabolic genotype-phenotype relationship. The series provides detailed coverage of the methodology for improving cellular properties of cells used in the production of biopharmaceuticals, gene and cell therapies and tissue engineering. It also seeks to explain the cellular mechanisms underlying in vitro physiological activity and productivity. This volume, which is based on presentations at the `European Workshop on Animal Cell Engineering' held in Costa Brava, Spain, contains a collection of chapters relating to cellular function and modification by leading authorities in several different areas of basic research and the biopharmaceutical industry.
Book Synopsis Cell Engineering by : Mohamed Al-Rubeai
Download or read book Cell Engineering written by Mohamed Al-Rubeai and published by Springer Science & Business Media. This book was released on 2007-11-23 with total page 311 pages. Available in PDF, EPUB and Kindle. Book excerpt: Integrating advances in molecular biology into bioprocesses presents a continuous challenge to scientists and bioengineers. This series is conceived to help meet this challenge. It examines and assesses the feasibility of new approaches for the modification of cellular function such as gene expression, protein processing, secretion, glycosylation, immortalisation, proliferation, and apoptosis as well as the systematic study of the metabolic genotype-phenotype relationship. The series provides detailed coverage of the methodology for improving cellular properties of cells used in the production of biopharmaceuticals, gene and cell therapies and tissue engineering. It also seeks to explain the cellular mechanisms underlying in vitro physiological activity and productivity. This volume, which is based on presentations at the `European Workshop on Animal Cell Engineering' held in Costa Brava, Spain, contains a collection of chapters relating to cellular function and modification by leading authorities in several different areas of basic research and the biopharmaceutical industry.
Current recombinant protein production systems require several months to develop. Existing systems fail to provide timely, flexible, and cost-effective therapies to protect against emergency mass-casualty infections or poisonings. As the identity of many new biological threat agents are unlikely to be known in advance, pre-emptive manufacturing and stockpiling of countermeasures cannot be performed. Preparedness for a biological catastrophe requires a radical solution to replace the current slow scale-up and manufacture of lifesaving medical countermeasures. Subunit vaccines and antibody fragments may be produced in bacteria, yeast, plant or insect cells. However, generation of full-length, human like glycosylated antibodies requires mammalian cell culture due to the cell’s ability to carry out complex assembly and processing. Although commercial cell culture methods for antibody production from stable gene expression have been substantially improved over the last 30 years, the time required to achieve full scale production for a new product is too long for a rapid, emergency response. An alternative method for rapid production of high quality antibody protein is transient gene expression. Transient gene expression is an established, routine approach to small scale, researchgrade material of recombinant proteins. It is frequently used to generate gram quantities of material within weeks of lead target identification. In the past, transient gene expression has been considered for emergency production of large production of antibodies, but dismissed due to low titers, high DNA requirements, uncertain regulations, unavailable manufacturing capacity, and uncertain scale-up performance. If these barriers can be overcome in the next few years, emergency use of transient gene expression for production of life saving medical countermeasures would be a viable means to help protect our nation from biological attacks. The goal of this thesis is to investigate both the technical and operational feasibility of scaling up transient gene expression. In order to investigate the technical feasibility of such a method, a phenomenological understanding of transfection was developed for process characterization, process optimization and scale-up studies. Experiments in shaker flasks and lab-scale bioreactors interrogated a number of factors involved in the transfection process and identified an optimal design space for performing transfections (Chapters 2 and 3). Important factors that were identified include cell/DNA/PEI ratio, transfection incubation time, and agitation set points. Through this optimization process, the highest reported titer (>300 mg/L) in transient CHO production was achieved. Experimental transfections also provided calibration metrics for phenomenological models of mass transfer in very large bioreactors. These models were used to investigate the potential of mass transfer limitations upon scale-up (Chapter 2). The results indicate that, with appropriate design of the agitation systems, including consideration of the impact of mass transfer of PEI/DNA complexes from the medium to the cells during the transfection stage, scale-up should be successful. In the final stage of experimentation, successful identification of scalable systems for aseptic liquid-cell separation eliminated other potential bottlenecks that may be encountered during scale-up (Chapter 3). A novel combination of existing technology generated simplified transfection protocols, which may be commercialized for alternative markets. Operational feasibility was investigated through a survey of current manufacturing capacity for mammalian cell culture and their capabilities to provide meaningful emergency production of antibody countermeasures. Process simulation was conducted to analyze process flow, plant design, and cost for large scale production of both plasmid DNA (Chapter 5), a sub-component of transient gene expression, as well as antibody protein (Chapter 4). Simulations predicted that a 1,000-L fermentor would produce sufficient plasmid DNA at a cost of approximately $377/gram. This DNA could be utilized in a 200,000-L facility to produce between 32 and 1,274 kg of recombinant antibody. Experimentally validated transfection processes were then used to refine simulations. Subsequent simulation resulted in production of 197 kg within 3 months at a cost of 705 $/gram mAb. These simulations predict that largescale transient gene expression can provide sufficient lifesaving countermeasures if titer improvements are possible and can be successfully scaled to large bioreactors. This thesis demonstrates both the operational and technical feasibility of a successful large-scale transient gene expression platform for the production of full-length, human-like glycosylated antibodies as medical countermeasures for biological catastrophe. This approach is one critical component of our Nation’s arsenal of bio-defense capabilities.
Book Synopsis Rapid, Large-Scale Production of Full-Length, Human-Like Monoclonal Antibodies by : Christopher M. Warner
Download or read book Rapid, Large-Scale Production of Full-Length, Human-Like Monoclonal Antibodies written by Christopher M. Warner and published by . This book was released on 2012 with total page 140 pages. Available in PDF, EPUB and Kindle. Book excerpt: Current recombinant protein production systems require several months to develop. Existing systems fail to provide timely, flexible, and cost-effective therapies to protect against emergency mass-casualty infections or poisonings. As the identity of many new biological threat agents are unlikely to be known in advance, pre-emptive manufacturing and stockpiling of countermeasures cannot be performed. Preparedness for a biological catastrophe requires a radical solution to replace the current slow scale-up and manufacture of lifesaving medical countermeasures. Subunit vaccines and antibody fragments may be produced in bacteria, yeast, plant or insect cells. However, generation of full-length, human like glycosylated antibodies requires mammalian cell culture due to the cell’s ability to carry out complex assembly and processing. Although commercial cell culture methods for antibody production from stable gene expression have been substantially improved over the last 30 years, the time required to achieve full scale production for a new product is too long for a rapid, emergency response. An alternative method for rapid production of high quality antibody protein is transient gene expression. Transient gene expression is an established, routine approach to small scale, researchgrade material of recombinant proteins. It is frequently used to generate gram quantities of material within weeks of lead target identification. In the past, transient gene expression has been considered for emergency production of large production of antibodies, but dismissed due to low titers, high DNA requirements, uncertain regulations, unavailable manufacturing capacity, and uncertain scale-up performance. If these barriers can be overcome in the next few years, emergency use of transient gene expression for production of life saving medical countermeasures would be a viable means to help protect our nation from biological attacks. The goal of this thesis is to investigate both the technical and operational feasibility of scaling up transient gene expression. In order to investigate the technical feasibility of such a method, a phenomenological understanding of transfection was developed for process characterization, process optimization and scale-up studies. Experiments in shaker flasks and lab-scale bioreactors interrogated a number of factors involved in the transfection process and identified an optimal design space for performing transfections (Chapters 2 and 3). Important factors that were identified include cell/DNA/PEI ratio, transfection incubation time, and agitation set points. Through this optimization process, the highest reported titer (>300 mg/L) in transient CHO production was achieved. Experimental transfections also provided calibration metrics for phenomenological models of mass transfer in very large bioreactors. These models were used to investigate the potential of mass transfer limitations upon scale-up (Chapter 2). The results indicate that, with appropriate design of the agitation systems, including consideration of the impact of mass transfer of PEI/DNA complexes from the medium to the cells during the transfection stage, scale-up should be successful. In the final stage of experimentation, successful identification of scalable systems for aseptic liquid-cell separation eliminated other potential bottlenecks that may be encountered during scale-up (Chapter 3). A novel combination of existing technology generated simplified transfection protocols, which may be commercialized for alternative markets. Operational feasibility was investigated through a survey of current manufacturing capacity for mammalian cell culture and their capabilities to provide meaningful emergency production of antibody countermeasures. Process simulation was conducted to analyze process flow, plant design, and cost for large scale production of both plasmid DNA (Chapter 5), a sub-component of transient gene expression, as well as antibody protein (Chapter 4). Simulations predicted that a 1,000-L fermentor would produce sufficient plasmid DNA at a cost of approximately $377/gram. This DNA could be utilized in a 200,000-L facility to produce between 32 and 1,274 kg of recombinant antibody. Experimentally validated transfection processes were then used to refine simulations. Subsequent simulation resulted in production of 197 kg within 3 months at a cost of 705 $/gram mAb. These simulations predict that largescale transient gene expression can provide sufficient lifesaving countermeasures if titer improvements are possible and can be successfully scaled to large bioreactors. This thesis demonstrates both the operational and technical feasibility of a successful large-scale transient gene expression platform for the production of full-length, human-like glycosylated antibodies as medical countermeasures for biological catastrophe. This approach is one critical component of our Nation’s arsenal of bio-defense capabilities.
This book represents the distillation and critical evaluation of many hundreds of publications relating to the production and use of antibodies. Therefore it is restricted to the "core" techniques of production and handling of antibodies, and their use in studies of antigen analysis, purification and localization.
Book Synopsis Monoclonal Antibodies by : James W. Goding
Download or read book Monoclonal Antibodies written by James W. Goding and published by . This book was released on 1986 with total page 344 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book represents the distillation and critical evaluation of many hundreds of publications relating to the production and use of antibodies. Therefore it is restricted to the "core" techniques of production and handling of antibodies, and their use in studies of antigen analysis, purification and localization.
Recombinant Protein Expression, Part B, Volume 660 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more. Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology serial Updated release includes the latest information on Recombinant Protein Expression
Book Synopsis Recombinant Protein Expression: Eukaryotic hosts by :
Download or read book Recombinant Protein Expression: Eukaryotic hosts written by and published by Academic Press. This book was released on 2021-11-04 with total page 384 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recombinant Protein Expression, Part B, Volume 660 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more. Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology serial Updated release includes the latest information on Recombinant Protein Expression
Engineered antibodies currently represent over 30% of biopharmaceuticals in clinical trials and their total worldwide sales continue to increase significantly. The importance of antibody applications is reflected in their increasing clinical and industrial applications as well as in the progression of established and emerging production strategies. This volume provides detailed coverage of the generation, optimization, characterization, production and applications of antibody. It provides the necessary theoretical background and description of methods for the expression of antibody in microbial and animal cell cultures and in transgenic animals and plants. There is a strong focus on those issues related to the production of intrabodies, bispecific antibody and antibody fragments and also to novel applications in cancer immunotherapy.
Book Synopsis Antibody Expression and Production by : Mohamed Al-Rubeai
Download or read book Antibody Expression and Production written by Mohamed Al-Rubeai and published by Springer. This book was released on 2011-05-18 with total page 346 pages. Available in PDF, EPUB and Kindle. Book excerpt: Engineered antibodies currently represent over 30% of biopharmaceuticals in clinical trials and their total worldwide sales continue to increase significantly. The importance of antibody applications is reflected in their increasing clinical and industrial applications as well as in the progression of established and emerging production strategies. This volume provides detailed coverage of the generation, optimization, characterization, production and applications of antibody. It provides the necessary theoretical background and description of methods for the expression of antibody in microbial and animal cell cultures and in transgenic animals and plants. There is a strong focus on those issues related to the production of intrabodies, bispecific antibody and antibody fragments and also to novel applications in cancer immunotherapy.
A valuable resource for researchers and workers in the fields of both pharmaceuticals and biotechnology as well as undergraduates in biochemistry, applied biology, biomedical sciences and pharmacy, this book compares established techniques of antibody production with the new. Antibody structure and the implications of antibody engineering are fully discussed, and a case study approach illustrates how antibodies are finding increasing use in the diagnosis and treatment of disease. The volume ends with commercial expression, purification and large-scale manufacture of antibodies and their future potential, particularly as therapeutic agents.
Book Synopsis Applications And Engineering Of Monoclonal Antibodies by : David J. King
Download or read book Applications And Engineering Of Monoclonal Antibodies written by David J. King and published by CRC Press. This book was released on 1998-11-27 with total page 262 pages. Available in PDF, EPUB and Kindle. Book excerpt: A valuable resource for researchers and workers in the fields of both pharmaceuticals and biotechnology as well as undergraduates in biochemistry, applied biology, biomedical sciences and pharmacy, this book compares established techniques of antibody production with the new. Antibody structure and the implications of antibody engineering are fully discussed, and a case study approach illustrates how antibodies are finding increasing use in the diagnosis and treatment of disease. The volume ends with commercial expression, purification and large-scale manufacture of antibodies and their future potential, particularly as therapeutic agents.
The Janeway's Immunobiology CD-ROM, Immunobiology Interactive, is included with each book, and can be purchased separately. It contains animations and videos with voiceover narration, as well as the figures from the text for presentation purposes.
Book Synopsis Janeway's Immunobiology by : Kenneth Murphy
Download or read book Janeway's Immunobiology written by Kenneth Murphy and published by Garland Science. This book was released on 2010-06-22 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: The Janeway's Immunobiology CD-ROM, Immunobiology Interactive, is included with each book, and can be purchased separately. It contains animations and videos with voiceover narration, as well as the figures from the text for presentation purposes.