Library Science

A core collection of diverse cutting-edge techniques for the generation, expression, optimization, and characterization of recombinant antibodies. Readily reproducible protocols for lead generation range from the cloning of human immunoglobulin genes to the selection and generation of human recombinant antibodies by humanization approaches, molecular display technologies and transgenic animals. Procedures are also described on restructuring antibody leads into monovalent, multivalent, and bispecific binding fragments for a wide variety of in vivo applications. State-of-the-art technologies are described for the characterization of antigen-binding affinity and specificity with novel applications in radioimmunotargeting, cancer immunotherapy, drug abuse, and proteomics.

Authors: Lo, Benny K. C.

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Table of Contents

1. Internet Resources for the Antibody Engineer (pp. 3-10)
   Lo, Benny K. C.; Chen, Yu Wai
2. The Kabat Database and a Bioinformatics Example (pp. 11-26)
   Johnson, George; Wu, Tai Te
3. IMGT, The International ImMunoGeneTics Information System®, http://imgt.cines.fr (pp. 27-50)
   Lefranc, Marie-Paule
4. Antibody Variable Regions: Toward a Unified Modeling Method (pp. 51-92)
   Whitelegg, Nicholas; Rees, Anthony R.
5. Studying Antibody Conformations by Ultracentrifugation and Hydrodynamic Modeling (pp. 93-114)
   Harding, Stephen E.; Longman, Emma; Carrasco, Beatriz; Ortega, Alvaro; de la Torre, Jose Garcia
6. PCR Cloning of Human Immunoglobulin Genes (pp. 117-134)
   Marks, James D.; Bradbury, Andrew
7. Antibody Humanization by CDR Grafting (pp. 135-160)
   Lo, Benny K. C.
8. Selection of Human Antibodies from Phage Display Libraries (pp. 161-176)
   Marks, James D.; Bradbury, Andrew
9. Production of Human Single-Chain Antibodies by Ribosome Display (pp. 177-190)
   He, Mingyue; Cooley, Neil; Jackson, Alison; J.Taussig, Michael
10. Production of Human Antibodies from Transgenic Mice (pp. 191-200)
    Davis, C. Geoffrey; Jia, Xiao-Chi; Feng, Xiao; Haak-Frendscho, Mary
11. Selection of Internalizing Antibodies for Drug Delivery (pp. 201-208)
    Marks, James D.
12. Engineering Multivalent Antibody Fragments for In Vivo Targeting (pp. 209-226)
    M.Wu, Anna
13. Production of Recombinant Bispecific Antibodies (pp. 227-242)
    Kontermann, Roland E.; Volkel, Tina; Korn, Tina
14. Expression and Isolation of Recombinant Antibody Fragments in E. coli (pp. 245-254)
    Charlton, Keith A.
15. Expression of Recombinant Antibodies in Mammalian Cell Lines (pp. 255-268)
    J.Yazaki, Paul; M.Wu, Anna
16. Human Antibody Production Using Insect-Cell Expression Systems (pp. 269-300)
    Guttieri, Mary C.; Liang, Mifang
17. Antibody Production in Transgenic Plants (pp. 301-318)
    Stoger, Eva; Schillberg, Stefan; M.Twyman, Richard; Fischer, Rainer; Christou, Paul
18. Directed Mutagenesis of Antibody Variable Domains (pp. 319-326)
    Brady, Kevin; Lo, Benny K. C.
19. Antibody Affinity Maturation by Chain Shuffling (pp. 327-344)
    Marks, James D.
20. Antibody Affinity Maturation by Random Mutagenesis (pp. 345-360)
    Fujii, Ikuo
21. Developing a Minimally Immunogenic Humanized Antibody by SDR Grafting (pp. 361-376)
    Kashmiri, Syed V. S.; De Pascalis, Roberto; Gonzales, Noreen R.
22. Antibody Purification by Column Chromatography (pp. 379-388)
    Murray, Andrea; Graves, C. Rosamund L.; Brady, Kevin; Lo, Benny K. C.
23. Affinity Measurement Using Surface Plasmon Resonance (pp. 389-416)
    Karlsson, Robert; Larsson, Anita
24. Kinetic Exclusion Assays to Study High-Affinity Binding Interactions in Homogeneous Solutions (pp. 417-430)
    Blake, Robert C.; Blake, Diane A.
25. Characterization of Antibody–Antigen Interactions by Fluorescence Spectroscopy (pp. 431-442)
    Missailidis, Sotiris; Brady, Kevin
26. Antibody Epitope Mapping Using Arrays of Synthetic Peptides (pp. 443-464)
    Reineke, Ulrich
27. Mapping Antibody:Antigen Interactions by Mass Spectrometry and Bioinformatics (pp. 465-480)
    Huhalov, Alexandra; Spencer, Daniel I.R.; Chester, Kerry A.
28. Radiometal Labeling of Antibodies and Antibody Fragments for Imaging and Therapy (pp. 481-494)
    Novak-Hofer, Ilse; Waibel, Robert; Zimmermann, Kurt; Schibli, Roger; Grunberg, Jurgen; Chester, Kerry A.; Murray, Andrea; Lo, Benny K.C.; Perkins, Alan C.; Schubiger, P. August
29. Production and Characterization of Anti-Cocaine Catalytic Antibodies (pp. 495-502)
    de Prada, Paloma; Landry, Donald W.
30. Recombinant Immunotoxins in the Treatment of Cancer (pp. 503-518)
    Pastan, Ira; Beers, Richard; Bera, Tapan K.
31. Antibodies in Proteomics (pp. 519-546)
    Bradbury, Andrew R. M.; Velappan, Nileena; Verzillo, Vittorio; Ovecka, Milan; Marzari, Roberto; Sblattero, Daniele; Chasteen, Leslie; Siegel, Robert; Pavlik, Peter
32. Targeting of Antibodies Using Aptamers (pp. 547-556)
    Missailidis, Sotiris

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