THE ANTIBODY MOLECULE

By JOSEPH MARTIN

CONTENTS OVERVIEW OF ANTIBODY PHAGE-DISPLAY TECHNOLOGY AND ITS APPLICATIONS STANDARD PROTOCOLS FOR THE CONSTRUCTION OF FAB LIBRARIES STANDARD PROTOCOLS FOR THE CONSTRUCTION OF SCFV LIBRARIES BROADENING THE IMPACT OF ANTIBODY PHAGE DISPLAY TECHNOLOGY: AMPLIFICATION OF IMMUNOGLOBULIN SEQUENCES FROM SPECIES OTHER THAN HUMANS OR MICE CONSTRUCTION OF LARGE NAÏVE FAB LIBRARIES CONSTRUCTION OF POLYCLONAL ANTIBODY LIBRARIES USING PHAGE DISPLAY ANTIGEN-DRIVEN STIMULATION OF B-LYMPHOCYTES IN VITRO THE RECOVERY OF IMMUNOGLOBULIN SEQUENCES FROM SINGLE HUMAN B CELLS BY CLONAL EXPANSION PANNING OF ANTIBODY PHAGE-DISPLAY LIBRARIES: STANDARD PROTOCOLS SELECTION OF ANTIBODIES AGAINST BIOTINYLATED ANTIGENS ISOLATION OF ANTI-HAPTEN SPECIFIC ANTIBODY FRAGMENTS FROM COMBINATORIAL LIBRARIES BLOCKING IMMUNODOMINANT EPITOPES BY COMPETITIVE DESELECTION SCREENING OF PHAGE-EXPRESSED ANTIBODY LIBRARIES BY CAPTURE LIFT ANTIBODY-GUIDED SELECTION USING CAPTURE-SANDWICH ELISA PROXIMITY-GUIDED (PROXIMOL) ANTIBODY SELECTION ISOLATION OF HUMAN MONOCLONAL ANTIBODIES USING GUIDED SELECTION WITH MOUSE MONOCLONAL ANTIBODIES SELECTING ANTIBODIES TO CELL-SURFACE ANTIGENS USING MAGNETIC SORTING TECHNIQUES ISOLATION OF HUMAN TUMOR-ASSOCIATED CELL SURFACE ANTIGEN-BINDING SCFVS SUBTRACTIVE ISOLATION OF SINGLE-CHAIN ANTIBODIES USING TISSUE FRAGMENTS SELECTION OF ANTIBODIES BASED ON ANTIBODY KINETIC BINDING PROPERTIES SELECTION OF FUNCTIONAL ANTIBODIES ON THE BASIS OF VALENCY TWO-STEP STRATEGY FOR ALTERATION OF IMMUNOGLOBULIN SPECIFICITY BY IN VITRO MUTAGENESIS TARGETING RANDOM MUTATIONS TO HOTSPOTS IN ANTIBODY VARIABLE DOMAINS FOR AFFINITY IMPROVEMENT ERROR-PRONE POLYMERASE CHAIN REACTION FOR MODIFICATION OF SCFVS USE OF ESCHERICHIA COLI MUTATOR CELLS TO MATURE ANTIBODIES CHAIN SHUFFLING TO MODIFY PROPERTIES OF RECOMBINANT IMMUNOGLOBULINS V ´ HIGH-LEVEL PERIPLASMIC EXPRESSION AND PURIFICATION OF SCFVS PERIPLASMIC EXPRESSION AND PURIFICATION OF RECOMBINANT FABS EXPRESSION OF ANTIBODY FRAGMENTS IN PICHIA PASTORIS EXPRESSION OF VHH ANTIBODY FRAGMENTS IN SACCHAROMYCES CEREVISIAE INTRABODIES: TARGETING SCFV EXPRESSION TO EUKARYOTIC INTRACELLULAR COMPARTMENTS EXPRESSION OF SCFVS AND SCFV FUSION PROTEINS IN EUKARYOTIC CELLS EXPRESSION OF ANTIBODY FAB FRAGMENTS AND WHOLE IMMUNOGLOBULIN IN MAMMALIAN CELLS