Antibody Engineering

Part I. Cloning of the Antigen-binding Site from Hybridoma

1. Cloning of Variable Domains from Mouse Hybridoma by PCR
Nina Strebe, Frank Breitling, Dieter Moosmayer, Bodo Brocks, Stefan Dübel

2. Coning Hybridoma cDNA by RACE
Andrew Bradbury

3. Construction of scFv Fragments from Hybridoma or Spleen Cells by PCR Assembly
Jonas V. Schaefer, Annemarie Honegger, Andreas Plückthun

Part II. Generation of Antibody Repertoires

4. Mouse Immune Libraries for the Generation of ScFv Fragments Directed Against Human Cell Surface Antigens
Christian Kellner, Sahar Mohseni Nodehi, Matthias Peipp

5. Human Antibody Gene Libraries
Michael Hust, Stefan Dübel

6. Synthetic Antibody Libraries
Pierre Martineau

7. Immune Libraries from Nonhuman Primates (NHP)
Thibaut Pelat, Michael Hust, Philippe Thullier

8. Generation of Rabbit Immune Libraries
Rüdiger Ridder, Hermann Gram

Part III. Selection of Antibody Fragments from Combinatorial Libraries

9. Immunotube Selections
Roland E. Kontermann

10. Phage Display and Selection in Microtitre Plates
Michael Hust, Michael Mersmann

11. Phage Display and Selections on Biotinylated Antigens
Patrick Chames, Daniel Baty

12. Phage Display and Subtractive Selection on Cells
Steffen U. Eisenhardt, Karlheinz Peter

13. Selection of Phage Antibody Libraries for Binding and Internalization into Mammalian Cells
Yu Zhou, James D. Marks

14. Improving Phage Display Throughput by Using Hyperphage, Miniaturized Titration and pVIII (g8p) ELISA
Frank Breitling, Olaf Broders, Saskia Helmsing, Michael Hust, Stefan Dübel

15. Yeast Display and Selections
Kelly Davis Orcutt, K. Dane Wittrup

16. The Generation of Transgenic Mice Expressing Human Antibody Repertoires
Xiangang Zou, Guocheng Lan, Michael J. Osborn, Marianne Brüggemann

17. Selection of Antibody Fragments by Means of the Filter-Sandwich Colony Screening Assay
Sabine Rauth, Martin Schlapschy, Arne Skerra

18. Semi-automated Magnetic Bead-Based Antibody Selection from Phage Display Libraries
Zoltán Konthur, Jeannine Wilde, Theam Soon Lim

Part IV. Engineering and Production of Immunoglobulins

19. Aspects of Isotype Selection
Zehra Kaymakcalan, Alexander Ibraghimov, Andrew G. Goodearl, Jochen G. Salfeld

20. Generation of Heavy and Light Chains (Chimeric Antibodies)
Kirstin A. Zettlitz

21. Humanising Antibodies by CDR Grafting
David Gareth Williams, David J. Matthews, Tarran Jones

22. Humanization by Resurfacing
Johan Desmet, Karen Vanhoorelbeke, Hans Deckmyn

23. Human Antibodies by Guided Selection
Sang Jick Kim, Insoo Park, Hyo Jeong Hong

24. In Silico De-Immunization
Philippe Alard, Johan Desmet, Ignace Lasters

25. Affinity Maturation by Chain Shuffling and Site Directed Mutagenesis
Jianlong Lou, James D. Marks

26. Affinity Maturation by Random Mutagenesis and Phage Display
Holger Thie

27. Engineering of the Fc Region for Improved PK (FcRn Interaction)
Vania E. Kenanova, Tove Olafsen, Jan T. Andersen, Inger Sandlie, Anna M. Wu

28. Antibody-Dependent Enzyme Prodrug Therapy (ADEPT)
Richard Begent, Surinder Sharma, Kerry Chester

29. Production of Recombinant Human IgG Antibodies in the Baculovirus Expression System
Mifang Liang, Stefan Dübel

30. Expression of IgA Molecules in Mammalian Cells
Thomas Beyer, Stefan Lohse, Michael Dechant, Thomas Valerius

Part V. Antibody Characterization

31. Expression of Complete Antibodies in Transgenic Plants
Doreen M. Floss, Udo Conrad

32. Expression of Full Length Monoclonal Antibodies (mAb) in Algal Chloroplast
Miller Tran, Stephen P. Mayfield

33. Expression of IgG Antibodies in Mammalian Cells
Thomas Jostock, Jiandong Li

34. Protein A/G Chromatography
Kirstin A. Zettlitz

35. Epitope Analysis Using Synthetic Peptide Repertoires Prepared by SPOT Synthesis Technology
U. Beutling, R. Frank

36. Epitope Mapping by Printed Peptide Libraries
Frank Breitling, Christopher Schirwitz, Thomas Felgenhauer, Ines Block, Volker Stadler, Ralf Bischoff

37. Antibody Epitope Mapping Using Yeast Display
Consuelo Garcia-Rodriguez, Yu Zhou, James D. Marks

38. Size Exclusion Chromatography
Bernd Voedisch, Holger Thie

39. Structural Characterization of Antibodies by Mass Spectrometry
Marie-Claire Janin-Bussat, Jean-Marc Strub, Elsa Wagner-Rousset, Olivier Colas, Christine Klinguer-Hamour, Nathalie Corvaia, Alain Dorsselaer, Alain Beck

40. Antibody Glycans Characterization
Marie-Claire Janin-Bussat, Elsa Wagner-Rousset, Christine Klinguer-Hamour, Nathalie Corvaia, Alain Dorsselaer, Alain Beck

41. Affinity Measurements by Competition ELISA
Pierre Martineau

42. Anti-Histidine Antibodies as Tools for Reversible Capturing of His-Tagged Fusion Proteins for Subsequent Binding Analysis
H.-M. Zenn, S. Hutschenreiter, F. W. Herberg

43. Affinity Measurements Using Quartz Crystal Microbalance (QCM)
Thomas Johansson

44. Affinity Measurements with Radiolabeled Antibodies
Verena Boschert, Peter Scheurich

45. Neutralization Tests
Philippe Thullier, Dorothea Thea Sesardic

46. Functional Characterization of Antibodies Neutralizing Soluble Factors In Vitro and In Vivo

Geertruida M. Veldman, Zehra Kaymakcalan, Renee Miller, Leena Kalghatgi, Jochen G. Salfeld

47. Competitive ELISA
Torsten Meyer

48. Quantification of Human IgG and Related Fc Fusion Proteins by a Human IgG/Fc Capture ELISA
Torsten Rülker, Doris Meier, Thomas Schirrmann

49. Determination of Fc-Mediated Antibody-Effector Functions by Chromium Release Assay
Tina Otz

50. Binding Studies with Flow Cytometry
Thomas Schirrmann

Nyckelord: Biomedicine, Antibodies, Protein Science, Cell Biology, Biological Techniques, Molecular Medicine