Monoclonal Antibodies : Meeting the Challenges in Manufacturing, Formulation, Delivery and Stability of Final Drug Product

Monoclonal Antibodies now have applications in virtually all areas of biology and medicine, and much of the world's biotechnology industry has its foundations in the exploitation of this technology. The Third Edition of this well established book meets the needs of both newcomers to the area and experienced researchers, by providing an integrated treatment of both the production and application of monoclonal antibodies. As in previous editions, detailed and critical accounts of the theory, production, purification, fragmentation, storage and radiolabelling of monoclonal antibodies are given, along with descriptions of their use in antigen characterization, affinity chromatography and immunofluorescence. The present volume has been comprehensively updated to cover recent rapid advances, particularly with respect to the applications of molecular biology, the use of antibodies in closing and heterologous expression of genes, immunohistology and phage display libraries. Since the previous edition, there has been a growing trend towards the replacement of procedures using radioactive isotopes, and the current edition incorporates these newer technologies. The text is oriented towards problems solving, and makes it easy to adapt each procedure to individual needs. Extensive cross-referencing, a glossary and a comprehensive index make this book an essential reference. This book will be vital both for laboratories already producing or using monoclonal antibodies, and for workers in many disciplines who are contemplating their use. * * Provides an integrated treatment of both the production and application of monoclonals in cell biology, biochemistry, and immunology. * Gives detailed and critical accounts of the theory, production, purification, storage, and relabelling of monoclonals, and their use in antigen characterization, affinity chromorography, and immunofluroscence. * Comprehensively updated to cover the rapid advances that have occurred since the publication of the Second Edition. Antigen Production For Monoclonal Antibody Generation / Giovanni Magistrelli And Pauline Malinge -- Method For The Generation Of Antibodies Specific For Site And Posttranslational Modifications / Hidemasa Goto And Masaki Inagaki -- Immunization, Hybridoma Generation, And Selection For Monoclonal Antibody Production / Hyung-yong Kim, Alexander Stojadinovic, And Mina J. Izadjoo -- Hybridoma Technology For The Generation Of Rodent Mabs Via Classical Fusion / Efthalia Chronopoulou ... [et Al.] -- Generation Of Rabbit Monoclonal Antibodies / Pi-chen Yam And Katherine L. Knight -- Screening Hybridomas For Cell Surface Antigens By High-throughput Homogeneous Assay And Flow Cytometry / Alejandro Uribe-benninghoff ... [et Al.] -- Screening And Subcloning Of High Producer Transfectomas Using Semisolid Media And Automated Colony Picker / Suba Dharshanan And Cheah Swee Hung -- Design And Generation Of Synthetic Antibody Libraries For Phage Display / Gang Chen And Sachdev S. Sidhu -- Selection And Screening Using Antibody Phage Display Libraries / Patrick Koenig And Germaine Fuh -- Yeast Surface Display For Antibody Isolation : Library Construction, Library Screening, And Affinity Maturation / James A.van Deventer And Karl Dane Wittrup -- Human B Cell Immortalization For Monoclonal Antibody Production / Joyce Hui-yuen, Siva Koganti, And Sumita Bhaduri-mcintosh -- Using Next-generation Sequencing For Discovery Of High- Frequency Monoclonal Antibodies In The Variable Gene Repertoires From Immunized Mice / Ulrike Haessler And Sai T. Reddy -- Cloning, Reformatting, And Small-scale Expression Of Monoclonal Antibody Isolated From Mouse, Rat, Or Hamster Hybridoma / Jeremy Loyau And François Rousseau -- Cloning Of Recombinant Monoclonal Antibodies From Hybridomas In A Single Mammalian Expression Plasmid / Nicole Müller-sienerth ... [et Al.] -- Monoclonal Antibody Purification By Ceramic Hydroxyapatite Chromatography / Larry J. Cummings, Russell G. Frost, And Mark A. Snyder -- Rapid Purification Of Monoclonal Antibodies Using Magnetic Microspheres / Pauline Malinge And Giovanni Magistrelli -- Generation Of Cell Lines For Monoclonal Antibody Production / Krista Alvin And Jianxin Ye -- Expression And Purification Of Recombinant Antibody Formats And Antibody Fusion Proteins / Martin Siegemund ... [et Al.] -- Purification Of Antibodies And Antibody Fragments Using Captureselect Affinity Resins / Pim Hermans, Hendrik Adams, And Frank Detmers -- Reformatting Of Scfv Antibodies Into The Scfv-fc Format And Their Downstream Purification / Emil Bujak, Mattia Matasci, Dario Neri, And Sarah Wulhfard -- Antibody V And C Domain Sequence, Structure, And Interaction Analysis With Special Reference To Imgt® / Eltaf Alamyar ... [et Al.] -- Measuring Antibody Affinities As Well As The Active Concentration Of Antigens Present On A Cell Surface / Palaniswami Rathanaswami -- Determination Of Antibody Structures / Robyn L. Stanfield -- Affinity Maturation Of Monoclonal Antibodies By Multi-site- Directed Mutagenesis / Hyung-yong Kim, Alexander Stojadinovic, And Mina J. Izadjoo -- Epitope Mapping With Membrane-bound Synthetic Overlapping Peptides / Terumi Midoro-horiuti And Randall M. Goldblum -- Epitope Mapping By Epitope Excision, Hydrogen/deuterium Exchange, And Peptide-panning Techniques Combined With In Silico Analysis/ Nicola Clementi ... [et Al.] -- Fine Epitope Mapping Based On Phage Display And Extensive Mutagenesis Of The Target Antigen / Gertrudis Rojas -- Epitope Mapping With Random Phage Display Library / Terumi Midoro-horiuti And Randall M. Goldblum -- Epitope Mapping Of Monoclonal And Polyclonal Antibodies Using Bacterial Cell Surface Display / Anna-luisa Volk, Francis Jingxin Hu, And Johan Rockberg -- Ion Exchange-high-performance Liquid Chromatography (iex-hplc) / Marie Corbier, Delphine Schrag, And Sylvain Raimondi -- Size Exclusion-high-performance Liquid Chromatography (sec-hplc) / Delphine Schrag, Marie Corbier, And Sylvain Raimondi -- N-glycosylation Characterization By Liquid Chromatography With Mass Spectrometry / Song Klapoetke -- Fc Engineering Of Antibodies And Antibody Derivatives By Primary Sequence Alteration And Their Functional Characterization / Stefanie Derer ... [et Al.] -- Labeling And Use Of Monoclonal Antibodies In Immunofluorescence : Protocols For Cytoskeletal And Nuclear Antigens / Christoph R. Bauer -- Generation And Use Of Antibody Fragments For Structural Studies Of Proteins Refractory To Crystallization / Stephen J. Stahl, Norman R. Watts, And Paul T. Wingfield -- Antibody Array Generation And Use / Carl A.k. Borrebaeck And Christer Wingren. Edited By Vincent Ossipow, Nicolas Fischer. Includes Bibliographical References And Index. Antigen production for monoclonal antibody generation -- Giovanni Magistrelli and Pauline Malinge Method for the generation of antibodies specific for site and posttranslational modifications -- Hidemasa Goto and Masaki Inagaki Immunization, hybridoma generation, and selection for monoclonal antibody production -- Hyung-Yong Kim, Alexander Stojadinovic, and Mina J. Izadjoo Hybridoma technology for the generation of rodent mAbs via classical fusion -- Efthalia Chronopoulou [and three others] Generation of rabbit monoclonal antibodies -- Pi-Chen Yam and Katherine L. Knight Screening hybridomas for cell surface antigens by high-throughput homogeneous assay and flow cytometry -- Alejandro Uribe-Benninghoff [and four others] Screening and subcloning of high producer transfectomas using semisolid media and automated colony picker -- Suba Dharshanan and Cheah Swee Hung Design and generation of synthetic antibody libraries for phage display -- Gang Chen and Sachdev S. Sidhu Selection and screening using antibody phage display libraries -- Patrick Koenig and Germaine Fuh Yeast surface display for antibody isolation : library construction, library screening, and affinity maturation -- James A.Van Deventer and Karl Dane Wittrup Human B cell immortalization for monoclonal antibody production -- Joyce Hui-Yuen, Siva Koganti, and Sumita Bhaduri-McIntosh Using next-generation sequencing for discovery of high- frequency monoclonal antibodies in the variable gene repertoires from immunized mice -- Ulrike Haessler and Sai T. Reddy Cloning, reformatting, and small-scale expression of monoclonal antibody isolated from mouse, rat, or hamster hybridoma -- Jeremy Loyau and François Rousseau Cloning of recombinant monoclonal antibodies from hybridomas in a single mammalian expression plasmid -- Nicole Müller-Sienerth [and three others] Monoclonal antibody purification by ceramic hydroxyapatite chromatography -- Larry J. Cummings, Russell G. Frost, and Mark A. Snyder Rapid purification of monoclonal antibodies using magnetic microspheres -- Pauline Malinge and Giovanni Magistrelli Generation of cell lines for monoclonal antibody production -- Krista Alvin and Jianxin Ye Expression and purification of recombinant antibody formats and antibody fusion proteins -- Martin Siegemund [and four others] Purification of antibodies and antibody fragments using CaptureSelect [trademark] affinity resins -- Pim Hermans, Hendrik Adams, and Frank Detmers Reformatting of scFv antibodies into the scFv-Fc format and their downstream purification -- Emil Bujak [and three others] Antibody V and C domain sequence, structure, and interaction analysis with special reference to IMGT® -- Eltaf Alamyar [and three others] Measuring antibody affinities as well as the active concentration of antigens present on a cell surface -- Palaniswami Rathanaswami Determination of antibody structures -- Robyn L. Stanfield Affinity maturation of monoclonal antibodies by multi-site- directed mutagenesis -- Hyung-Yong Kim, Alexander Stojadinovic, and Mina J. Izadjoo Epitope mapping with membrane-bound synthetic overlapping peptides -- Terumi Midoro-Horiuti and Randall M. Goldblum Epitope mapping by epitope excision, hydrogen/deuterium exchange, and peptide-panning techniques combined with in silico analysis -- Nicola Clementi [and five others] Fine epitope mapping based on phage display and extensive mutagenesis of the target antigen -- Gertrudis Rojas Epitope mapping with random phage display library -- Terumi Midoro-Horiuti and Randall M. Goldblum Epitope mapping of monoclonal and polyclonal antibodies using bacterial cell surface display -- Anna-Luisa Volk, Francis Jingxin Hu, and Johan Rockberg Ion exchange-high-performance liquid chromatography (IEX-HPLC) -- Marie Corbier, Delphine Schrag, and Sylvain Raimondi Size exclusion-high-performance liquid chromatography (SEC-HPLC) -- Delphine Schrag, Marie Corbier, and Sylvain Raimondi N-glycosylation characterization by liquid chromatography with mass spectrometry -- Song Klapoetke Fc engineering of antibodies and antibody derivatives by primary sequence alteration and their functional characterization -- Stefanie Derer [and six others] Labeling and use of monoclonal antibodies in immunofluorescence : protocols for cytoskeletal and nuclear antigens -- Christoph R. Bauer Generation and use of antibody fragments for structural studies of proteins refractory to crystallization -- Stephen J. Stahl, Norman R. Watts, and Paul T. Wingfield Antibody array generation and use -- Carl A.K. Borrebaeck and Christer Wingren. Annotation. Monoclonal Antibodies now have applications in virtually all areas of biology and medicine, and much of the world's biotechnology industry has its foundations in the exploitation of this technology. The Third Edition of this well established book meets the needs of both newcomers to the area and experienced researchers, by providing an integrated treatment of both the production and application of monoclonal antibodies. As in previous editions, detailed and critical accounts of the theory, production, purification, fragmentation, storage and radiolabelling of monoclonal antibodies are given, along with descriptions of their use in antigen characterization, affinity chromatography and immunofluorescence. The present volume has been comprehensively updated to cover recent rapid advances, particularly with respect to the applications of molecular biology, the use of antibodies in closing and heterologous expression of genes, immunohistology and phage display libraries. Since the previous edition, there has been a growing trend towards the replacement of procedures using radioactive isotopes, and the current edition incorporates these newer technologies. The text is oriented towards problems solving, and makes it easy to adapt each procedure to individual needs. Extensive cross-referencing, a glossary and a comprehensive index make this book an essential reference. This book will be vital both for laboratories already producing or using monoclonal antibodies, and for workers in many disciplines who are contemplating their use. * * Provides an integrated treatment of both the production and application of monoclonals in cell biology, biochemistry, and immunology. * Gives detailed and critical accounts of the theory, production, purification, storage, and relabelling of monoclonals, and their use in antigen characterization, affinity chromorography, and immunofluroscence. * Comprehensively updated to cover the rapid advances that have occurred since the publication of the Second Edition.

Monoclonal antibodies (MAbs) are currently the major class of protein bio therapeutic being developed by biotechnology and pharmaceutical companies. Monoclonal Antibodies discusses the challenges and issues revolving around development of a monoclonal antibody produced by recombinant DNA technology into a therapeutic agent.This book covers downstream processing which includes design of processes to manufacture the formulation, formulation design, fill and finish into closure systems and routes of administration. The characterization of the final drug product is covered where the use of biophysical methods combined with genetic engineering is used to understand the solution properties of the formulation. The latter has become very important since many indications such as arthritis and asthma require the development of formulations for subcutaneous delivery (SC). The development of formulations for IV delivery is also important and comes with a different set of challenges. The challenges and strategies that can overcome these limitations are discussed in this book, starting with an introduction to these issues, followed by chapters detailing strategies to deal with them. Subsequent chapters explore the processing and storage of mAbs, development of delivery device technologies and conclude with a chapter on the future of mAbs in therapeutic remedies.

• Discusses the challenges to develop MAbs for intravenous (IV) and subcutaneous delivery (SC)
• Presents strategies to meet the challenges in development of MAbs for SC and IV administration
• Discusses the use of biophysical analytical tools coupled with MAb engineering to understand what governs MAb properties at high concentration

Monoclonal antibodies (MAbs) are currently the major class of protein bio therapeutic being developed by biotechnology and pharmaceutical companies. Monoclonal Antibodies discusses the challenges and issues revolving around development of a monoclonal antibody produced by recombinant DNA technology into a therapeutic agent.This book covers downstream processing which includes design of processes to manufacture the formulation, formulation design, fill and finish into closure systems and routes of administration. The characterization of the final drug product is covered where the use of biophysical methods combined with genetic engineering is used to understand the solution properties of the formulation. The latter has become very important since many indications such as arthritis and asthma require the development of formulations for subcutaneous delivery (SC). The development of formulations for IV delivery is also important and comes with a different set of challenges. The challenges and strategies that can overcome these limitations are discussed in this book, starting with an introduction to these issues, followed by chapters detailing strategies to deal with them. Subsequent chapters explore the processing and storage of mAbs, development of delivery device technologies and conclude with a chapter on the future of mAbs in therapeutic remedies. Discusses the challenges to develop MAbs for intravenous (IV) and subcutaneous delivery (SC) Presents strategies to meet the challenges in development of MAbs for SC and IV administration Discusses the use of biophysical analytical tools coupled with MAb engineering to understand what governs MAb properties at high concentration Monoclonal antibodies (MAbs) are currently the major class of protein bio therapeutic being developed by biotechnology and pharmaceutical companies. Monoclonal Antibodies discusses the challenges and issues revolving around development of a monoclonal antibody produced by recombinant DNA technology into a therapeutic agent. This book covers downstream processing which includes design of processes to manufacture the formulation, formulation design, fill and finish into closure systems and routes of administration. The characterization of the final drug product is covered where the use of biophysical methods combined with genetic engineering is used to understand the solution properties of the formulation. The latter has become very important since many indications such as arthritis and asthma require the development of formulations for subcutaneous delivery (SC). The development of formulations for IV delivery is also important and comes with a different set of challenges. The challenges and strategies that can overcome these limitations are discussed in this book, starting with an introduction to these issues, followed by chapters detailing strategies to deal with them. Subsequent chapters explore the processing and storage of mAbs, development of delivery device technologies and conclude with a chapter on the future of mAbs in therapeutic remedies. Discusses the challenges to develop MAbs for intravenous (IV) and subcutaneous delivery (SC) Presents strategies to meet the challenges in development of MAbs for SC and IV administration Discusses the use of biophysical analytical tools coupled with MAb engineering to understand what governs MAb properties at high concentration Monoclonal Antibodies: Methods and Protocols, Second Edition expands upon the previous edition with current, detailed modern approaches to isolate and characterize monoclonal antibodies against carefully selected epitopes. This edition includes new chapters covering the key steps to generate high quality monoclonals via different methods, from antigen generation to epitope mapping and quality control of the purified IgG. Chapters are divided into four parts corresponding to four distinct objectives. Part I covers monoclonal antibody generation, Part II deals with monoclonal antibody expression and purification, Part III presents methods for monoclonal antibody characterization and modification, and Part IV describes selected applications of monoclonal antibodies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Monoclonal Antibodies: Methods and Protocols, Second Edition provides crucial initial steps of monoclonal antibody generation and characterization with state-of-the art protocols.-- Front Cover; Related titles; Monoclonal Antibodies; Copyright; Contents; List of figures; List of tables; About the author; Preface; 1 - Introduction; Pharmaceutical development; Development of the API; mAbs as protein therapeutics; Brief review of mAb structure; References; 2 - Analytical tools used in the formulation and assessment of stability of monoclonal antibodies (mAbs); Analytical methods for evaluation of monoclonal antibody stability; References; 3 - Stability of monoclonal antibodies (mAbs); Degradation routes in monoclonal antibodies; Chemical degradation; Mechanisms of oxidation This edition reflects the advances that have occured in the field of monoclonal antibodies since the mid-1980s. It provides an integrated treatment of both the production and application of monoclonals in cell biology, biochemistry and immunology