Antibody Engineering

1 Recombinant Antibodies.- Generation of Recombinant Antibodies from Hybridoma.- 2 Construction of scFv Fragments from Hybridoma or Spleen Cells by PCR Assembly.- 3 Construction of scFv from Hybridoma by Two-Step Cloning.- 4 Cloning Hybridoma cDNA by RACE.- Generation of Antibody Libraries.- 5 Phage-Display Libraries of Murine Antibody Fab Fragments.- 6 Generation of Naive Human Antibody Libraries.- 7 Generation of Antibody Gene Libraries from Seropositive Human Donors.- 8 Generation of Rabbit Immune Libraries.- Isolation of Antibody Fragments from Combinatorial Libraries.- 9 Immunotube Selections.- 10 Selections on Biotinylated Antigens.- 11 Guided Selections on Cells.- 12 Proximity (ProxiMol) and Step-Back Selections.- 13 Selections on Tissue Sections.- 14 Selections of Human Antibody Fragments by Anti-Idiotypic Interaction.- 15 Selecting Intracellular Antibodies Using the Two-Hybrid System.- 16 Selection of Phage Antibody Libraries for Binding and Internalization Into Mammalian Cells.- Expression and Purification of Antibody Fragments in Bacteria.- 17 Purification and Characterisation of His-Tagged Antibody Fragments.- 18 Functional Minipreps of scFv-Antibody Fragments.- 19 Expression of scFv in L-Form Bacteria.- 20 Expression and Purification of Fab Fragments.- 21 Purification and Analysis of Antibody Fragments Using Proteins L, A and LA.- 22 Purification and Analysis of Strep-Tagged Antibody Fragments.- 23 Improving Expression of scFv Fragments by Coexpression of Periplasmic Chaperons.- Expression and Purification of Antibody Fragments in Eukaryotic Cells.- 24 Expresssion of scFv Antibody Fragments in the Yeast Pichia Pastoris.- 25 Production of Recombinant Human IgG Antibodies in the Baculovirus Expresssion System.- 26 Expresssion of Antibodies in Mammalian Cells.- 27 Expresssion of Antibody Fragments in Plant Cells.- Determination of Affinities.- 28 Affinity Measurements by Band Shift and Competition ELISA.- 29 Affinity Measurements of Antibody Fragments on Phage by Quartz Crystal Microbalance (QCM).- Sequence and Structure Analysis and Modelling.- 30 Sequence Analysis of Human Antibody Genes.- 31 Protein Sequence and Structure Analysis of Antibody Variable Domains.- Epitope Mapping.- 32 Epitope Mapping with Synthetic Peptides Prepared by SPOT Synthesis.- 33 Epitope Mapping with Random Peptide Libraries.- 34 Epitope Mapping with Gene Fragment Libraries.- In vivo Experiments.- 35 Imaging Tumor Xenografts Using Radiolabeled Antibody Fragments.- 36 Xenograft Mouse Models for Tumour Targeting.- Affinity Maturation of Antibody Fragments.- 37 Affinity Maturation by Chain Shuffling and Site Directed Mutagenesis.- 38 Affinity and Specificity Maturation by CDR Walking.- Humanisation of Antibody Fragments.- 39 Humanisation by Guided Selection.- 40 Humanising Antibodies by CDR Grafting.- Antibody Engineering to Improve Stability.- 41 Stabilization Strategies and Application of recombinant Fvs and Fv Fusion Proteins.- Bivalent and Bispecific Antibody Fragments.- 42 Bivalent and Bispecific Diabodies and Single-Chain Diabodies.- 43 Miniantibodies.- 44 ScFv-CH3 and scFv-Fc Fusion Proteins.- 45 CTL-Assays for Functional Testing of Bispecific Antibody Fragments.- Recombinant Antibody-Fusion Proteins.- 46 Antibody-Enzyme Fusions.- 47 Cytokine-scFv Fusion Proteins.- 48 Antibody Fusion Proteins for Targeted Gene Delivery.- Intracellular Targeting of Antibody Fragments.- 49 Intracellular Single Chain Antibodies - Methods for Derivation and Employment.- 50 Expresssing Intracellular Single-Chain Fv Fragments in Mammalian Cells.- Appendices.- Appendix 1 Internet Resources.- Appendix 2 Amino Acids: Nomenclature and Codons.