1;Foreword;5 2;Preface;6 3;Contents;7 4;Cloning of the Antigen-binding Site from Hybridoma;12 4.1;Cloning of Variable Domains from Mouse Hybridoma by PCR;13 4.2;Coning Hybridoma cDNA by RACE;25 4.3;Construction of scFv Fragments from Hybridoma or Spleen Cells by PCR Assembly;31 5;Generation of Antibody Repertoires;55 5.1;Mouse Immune Libraries for the Generation of ScFv Fragments Directed Against Human Cell Surface Antigens;56 5.2;Human Antibody Gene Libraries;73 5.3;Synthetic Antibody Libraries;93 5.4;Immune Libraries from Nonhuman Primates ( NHP);106 5.5;Generation of Rabbit Immune Libraries;122 6;Selection of Antibody Fragments from Combinatorial Libraries;131 6.1;Immunotube Selections;132 6.2;Phage Display and Selection in Microtitre Plates;143 6.3;Phage Display and Selections on Biotinylated Antigens;154 6.4;Phage Display and Subtractive Selection on Cells;168 6.5;Selection of Phage Antibody Libraries for Binding and Internalization into Mammalian Cells;186 6.6;Improving Phage Display Throughput by Using Hyperphage, Miniaturized Titration and pVIII ( g8p) ELISA;199 6.7;Yeast Display and Selections;209 6.8;The Generation of Transgenic Mice Expressing Human Antibody Repertoires;236 6.9;Selection of Antibody Fragments by Means of the Filter- Sandwich Colony Screening Assay;256 6.10;Semi-automated Magnetic Bead-Based Antibody Selection from Phage Display Libraries;268 7;Engineering and Production of Immunoglobulins;289 7.1;Aspects of Isotype Selection;290 7.2;Generation of Heavy and Light Chains ( Chimeric Antibodies);306 7.3;Humanising Antibodies by CDR Grafting;317 7.4;Humanization by Resurfacing;338 7.5;Human Antibodies by Guided Selection;351 7.6;In Silico De-Immunization;365 7.7;Affinity Maturation by Chain Shuffling and Site Directed Mutagenesis;373 7.8;Affinity Maturation by Random Mutagenesis and Phage Display;393 7.9;Engineering of the Fc Region for Improved PK ( FcRn Interaction);406 7.10;Antibody-Dependent Enzyme Prodrug Therapy ( ADEPT);426 7.11;Production of Recombinant Human IgG Antibodies in the Baculovirus Expression System;447 7.12;Expression of IgA Molecules in Mammalian Cells;465 8;Antibody Characterization;481 8.1;Expression of Complete Antibodies in Transgenic Plants;482 8.2;Expression of Full Length Monoclonal Antibodies ( mAb) in Algal Chloroplast;496 8.3;Expression of IgG Antibodies in Mammalian Cells;510 8.4;Protein A/G Chromatography;523 8.5;Epitope Analysis Using Synthetic Peptide Repertoires Prepared by SPOT Synthesis Technology;528 8.6;Epitope Mapping by Printed Peptide Libraries;563 8.7;Antibody Epitope Mapping Using Yeast Display;580 8.8;Size Exclusion Chromatography;595 8.9;Structural Characterization of Antibodies by Mass Spectrometry;601 8.10;Antibody Glycans Characterization;623 8.11;Affinity Measurements by Competition ELISA;645 8.12;Anti-Histidine Antibodies as Tools for Reversible Capturing of His- Tagged Fusion Proteins for Subsequent Binding Analysis;654 8.13;Affinity Measurements Using Quartz Crystal Microbalance ( QCM);669 8.14;Affinity Measurements with Radiolabeled Antibodies;680 8.15;Neutralization Tests;690 8.16;Functional Characterization of Antibodies Neutralizing Soluble Factors In Vitro and In Vivo;708 8.17;Competitive ELISA;723 8.18;Quantification of Human IgG and Related Fc Fusion Proteins by a Human IgG/ Fc Capture ELISA;727 8.19;Determination of Fc-Mediated Antibody- Effector Functions by Chromium Release Assay;733 8.20;Binding Studies with Flow Cytometry;748 9;Index;763