Antibody Phage Display: Methods and Protocols

Contents......Page 7
1 Overview of Antibody Phage-Display Technology and Its Applications......Page 13
2 Standard Protocols for the Construction of Fab Libraries......Page 50
3 Standard Protocols for the Construction of scFv Libraries......Page 70
4 Broadening the Impact of Antibody Phage Display Technology:......Page 83
5 Construction of Large Naïve Fab Libraries......Page 97
6 Construction of Polyclonal Antibody Libraries Using Phage Display......Page 111
7 Antigen-Driven Stimulation of B-Lymphocytes In Vitro......Page 123
8 The Recovery of Immunoglobulin Sequences from Single Human......Page 130
9 Panning of Antibody Phage-Display Libraries: Standard Protocols......Page 141
10 Selection of Antibodies Against Biotinylated Antigens......Page 154
11 Isolation of Anti-Hapten Specific Antibody Fragments......Page 165
12 Blocking Immunodominant Epitopes by Competitive Deselection......Page 178
13 Rescue of a Broader Range of Antibody Specificities Using......Page 183
14 Screening of Phage-Expressed Antibody Libraries by Capture Lift......Page 191
15 Antibody-Guided Selection Using Capture-Sandwich ELISA......Page 198
16 Proximity-Guided (ProxiMol) Antibody Selection......Page 203
17 Isolation of Human Monoclonal Antibodies Using Guided Selection
with Mouse Monoclonal Antibodies......Page 208
18 Selecting Antibodies to Cell-Surface Antigens Using Magnetic
Sorting Techniques......Page 219
19 Isolation of Human Tumor-Associated Cell Surface
Antigen-Binding scFvs......Page 227
20 Subtractive Isolation of Single-Chain Antibodies Using
Tissue Fragments......Page 234
21 Selection of Antibodies Based on Antibody Kinetic Binding Properties
v
′......Page 243
22 Selection of Functional Antibodies on the Basis of Valency......Page 252
23 Two-Step Strategy for Alteration of Immunoglobulin Specificity
by In Vitro Mutagenesis......Page 256
24 Targeting Random Mutations to Hotspots in Antibody Variable
Domains for Affinity Improvement......Page 265
25 Error-Prone Polymerase Chain Reaction for Modification of scFvs......Page 282
26 Use of Escherichia coli Mutator Cells to Mature Antibodies......Page 290
27 Chain Shuffling to Modify Properties of Recombinant
Immunoglobulins......Page 298
28 Generation of Bispecific and Tandem Diabodies......Page 312
29 High-Level Periplasmic Expression and Purification of scFvs......Page 327
30 Periplasmic Expression and Purification of Recombinant Fabs......Page 336
31 Expression of Antibody Fragments in Pichia pastoris......Page 342
32 Expression of VHH Antibody Fragments in Saccharomyces cerevisiae......Page 351
33 Intrabodies: Targeting scFv Expression to Eukaryotic Intracellular
Compartments......Page 359
34 Expression of scFvs and scFv Fusion Proteins in Eukaryotic Cells......Page 371
Index......Page 387
guoxingzhong and huangzhiman......Page 2
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