Antibody Engineering Volume 2 (eBook)

Foreword 5
Preface 6
Contents 7
Bioinformatics of Antigen-binding Sites 11
Analysis of Single Chain Antibody Sequences Using the VBASE2 Fab Analysis Tool 12
Standardized Sequence and Structure Analysis of Antibody Using IMGT1 20
Protein Sequence and Structure Analysis of Antibody Variable Domains 41
Generation of Antibody Fragments and Their Derivatives 60
scFv by Two-Step Cloning 61
Bivalent Diabodies 66
Generation of Single-Chain Fv Fragments and Multivalent Derivatives scFv- Fc and scFv- CH3 ( Minibodies) 74
Miniantibodies 90
Generation of Stably Transfected Eukaryotic Cell Lines Producing ImmunoRNAse Fusion Proteins 105
Antibody–Cytokine Fusion Proteins with Members of the TNF- Family 117
Recombinant Immunotoxins for Treating Cancer 131
T Bodies 151
Expressing Intrabodies in Mammalian Cells 164
Phenotypic Knockdown with Intrabodies 176
Disulfide-Stabilized Fv Fragments 184
PEGylation of Antibody Fragments to Improve Pharmacodynamics and Pharmacokinetics 193
Fusion Proteins with Improved PK 208
In Vivo Biotinylated scFv Fragments 220
Bispecific Diabodies and Single-Chain Diabodies 228
Generation and Characterization of a Dual Variable Domain Immunoglobulin Molecule 239
Isolation of Antigen-Specific Nanobodies 251
CDR-FR Peptides 267
Production of Antibody Fragments 277
Purification and Characterization of His-Tagged Antibody Fragments 278
Production of Antibody Fragments in the Gram- Positive Bacterium Bacillus megaterium 291
Analysis and Purification of Antibody Fragments Using Protein A, Protein G, and Protein L 298
Purification and Analysis of Strep-tagged Antibody- Fragments 313
Production of Antibodies and Antibody Fragments in Escherichia coli 327
Improving Expression of scFv Fragments by Co- expression of Periplasmic Chaperones 341
Bioreactor Production of scFv Fragments in Pichia pastoris 358
Expression of Antibody Fragments in Transgenic Plants 371
Transient Production of scFv-Fc Fusion Proteins in Mammalian Cells 381
Recombinant Antibody Molecules in Nanobiotechnology and Proteomics 393
Immunoliposomes 394
Targeted Polymeric Nanoparticles 410
Antibody Microarrays for Expression Analysis 422
Evaluation of Recombinant Antibodies on Protein Microarrays Applying the Multiple Spotting Technique 439
Preclinical and Clinical Development 453
Xenograft Mouse Models for Tumour Targeting 454
Xenograft Mouse Models for Tumour Targeting 468
Imaging Tumor Xenografts Using Radiolabeled Antibodies 482
Human Anti-antibody Response 498
IP Issues in the Therapeutic Antibody Industry 507
Appendix 572
Index 574