Main description:

The rapidly growing field of antibody research is the result of many advancing technologies allowing current developments to take advantage of molecular engineering to create tailor-made antibodies. Antibody Methods and Protocols attempts to provide insight into the generation of antibodies using in vitro and in vivo approaches, as well as technical aspects for screening, analysis, and modification of antibodies and antibody fragments. The detailed volume is focused on basic protocols for isolating antibodies and, at the same time, it selects a range of specific areas with the aim of providing guides for the overall process of antibody isolation and characterization as well as protocols for enhancing classical antibodies and antibody fragments. Written in the highly successful Methods in Molecular Biology (TM) 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 easy to use, Antibody Methods and Protocols provides a broad and useful background to support ongoing efforts by novices and experts alike and encourages the development of new imaginative approaches to this vital area of study.

Contents:

1. Antigen Presentation for the Generation of Binding Molecules

Hilmar Ebersbach, Gabriele Proetzel, and Chonghui Zhang

2. Recombinant Antibodies and In Vitro Selection Technologies

C. Ronald Geyer, John McCafferty, Stefan Dubel, Andrew R.M. Bradbury, and Sachdev S. Sidhu

3. Phage Display

Konstantin Petropoulos

4. Generation of Human Fab Libraries for Phage Display

Christoph Rader

5. Selection of Human Fab Libraries by Phage Display

Christoph Rader

6. Ribosome Display

George Thom and Maria Groves

7. Hybridoma Technology for the Generation of Monoclonal Antibodies

Chonghui Zhang

8. The Application of Transgenic Mice for Therapeutic Antibody Discovery

E-Chiang Lee and Michael Owen

9. Production of Human or Humanized Antibodies in Mice

Brice Laffleur, Virginie Pascal, Christophe Sirac, and Michel Cogne

10. Immortalization of Human B Cells: Analysis of B Cell Repertoire and Production of Human Monoclonal Antibodies

Elisabetta Traggiai

11. Kinetic Screening in the Antibody Development Process

Michael Schraml and Matthias Biehl

12. Temperature-Dependent Antibody Kinetics as a Tool in Antibody Lead Selection

Michael Schraml and Leopold von Proff

13. Determination of Antibody Glycosylation by Mass Spectrometry

Christiane Jager, Claudia Ferrara, Pablo Umana, Anne Zeck, Joerg Thomas Regula, and Hans Koll

14. Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format

Jiahui Yang and Christoph Rader

15. PEGylation of Antibody Fragments for Half-Life Extension

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Konstantin Petropoulos

4. Generation of Human Fab Libraries for Phage Display

Christoph Rader

5. Selection of Human Fab Libraries by Phage Display

Christoph Rader

6. Ribosome Display

George Thom and Maria Groves

7. Hybridoma Technology for the Generation of Monoclonal Antibodies

Chonghui Zhang

8. The Application of Transgenic Mice for Therapeutic Antibody Discovery

E-Chiang Lee and Michael Owen

9. Production of Human or Humanized Antibodies in Mice

Brice Laffleur, Virginie Pascal, Christophe Sirac, and Michel Cogne

10. Immortalization of Human B Cells: Analysis of B Cell Repertoire and Production of Human Monoclonal Antibodies

Elisabetta Traggiai

11. Kinetic Screening in the Antibody Development Process

Michael Schraml and Matthias Biehl

12. Temperature-Dependent Antibody Kinetics as a Tool in Antibody Lead Selection

Michael Schraml and Leopold von Proff

13. Determination of Antibody Glycosylation by Mass Spectrometry

Christiane Jager, Claudia Ferrara, Pablo Umana, Anne Zeck, Joerg Thomas Regula, and Hans Koll

14. Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format

Jiahui Yang and Christoph Rader

15. PEGylation of Antibody Fragments for Half-Life Extension

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Brice Laffleur, Virginie Pascal, Christophe Sirac, and Michel Cogne

10. Immortalization of Human B Cells: Analysis of B Cell Repertoire and Production of Human Monoclonal Antibodies

Elisabetta Traggiai

11. Kinetic Screening in the Antibody Development Process

Michael Schraml and Matthias Biehl

12. Temperature-Dependent Antibody Kinetics as a Tool in Antibody Lead Selection

Michael Schraml and Leopold von Proff

13. Determination of Antibody Glycosylation by Mass Spectrometry

Christiane Jager, Claudia Ferrara, Pablo Umana, Anne Zeck, Joerg Thomas Regula, and Hans Koll

14. Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format

Jiahui Yang and Christoph Rader

15. PEGylation of Antibody Fragments for Half-Life Extension

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Elisabetta Traggiai

11. Kinetic Screening in the Antibody Development Process

Michael Schraml and Matthias Biehl

12. Temperature-Dependent Antibody Kinetics as a Tool in Antibody Lead Selection

Michael Schraml and Leopold von Proff

13. Determination of Antibody Glycosylation by Mass Spectrometry

Christiane Jager, Claudia Ferrara, Pablo Umana, Anne Zeck, Joerg Thomas Regula, and Hans Koll

14. Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format

Jiahui Yang and Christoph Rader

15. PEGylation of Antibody Fragments for Half-Life Extension

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Michael Schraml and Matthias Biehl

12. Temperature-Dependent Antibody Kinetics as a Tool in Antibody Lead Selection

Michael Schraml and Leopold von Proff

13. Determination of Antibody Glycosylation by Mass Spectrometry

Christiane Jager, Claudia Ferrara, Pablo Umana, Anne Zeck, Joerg Thomas Regula, and Hans Koll

14. Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format

Jiahui Yang and Christoph Rader

15. PEGylation of Antibody Fragments for Half-Life Extension

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Michael Schraml and Leopold von Proff

13. Determination of Antibody Glycosylation by Mass Spectrometry

Christiane Jager, Claudia Ferrara, Pablo Umana, Anne Zeck, Joerg Thomas Regula, and Hans Koll

14. Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format

Jiahui Yang and Christoph Rader

15. PEGylation of Antibody Fragments for Half-Life Extension

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

13. Determination of Antibody Glycosylation by Mass Spectrometry

Christiane Jager, Claudia Ferrara, Pablo Umana, Anne Zeck, Joerg Thomas Regula, and Hans Koll

14. Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format

Jiahui Yang and Christoph Rader

15. PEGylation of Antibody Fragments for Half-Life Extension

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Christiane Jager, Claudia Ferrara, Pablo Umana, Anne Zeck, Joerg Thomas Regula, and Hans Koll

14. Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format

Jiahui Yang and Christoph Rader

15. PEGylation of Antibody Fragments for Half-Life Extension

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Jiahui Yang and Christoph Rader

15. PEGylation of Antibody Fragments for Half-Life Extension

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Simona Jevsevar, Mateja Kusterle, and Maja Kenig

16. Bispecific Antibody Derivatives Based on Full-Length IgG Formats

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Michael Grote, Alexander K. Haas, Christian Klein, Wolfgang Schaefer, and Ulrich Brinkmann

17. Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Alexander K. Haas, Klaus Mayer, and Ulrich Brinkmann

18. Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Robert F. Kelley and Y. Gloria Meng

19. Class-Specific Effector Functions of Therapeutic Antibodies

Virginie Pascal, Brice Laffleur, and Michel Cogne

Virginie Pascal, Brice Laffleur, and Michel Cogne