Selection of an anti-IGF-1 Fab from a Fab phage library created by mutagenesis of multiple CDR loops.
Diverse Fab libraries containing 2-3 x 10(8) members were generated by randomizing amino acid residues within four of the six complementarity determining regions of a humanized version of an anti-HER-2 Ab (hu4D5). These libraries were subsequently displayed on the surface of the filamentous bacteriophage M13 and selected for binding to three proteins: CD4, insulin-like growth factor 1 (IGF-1), and tissue plasminogen activator. An Fab-bacteriophage was isolated that showed specific binding to IGF-1. The affinity of this Fab was determined to be 3.5 microM.
Garrard
L J
LJ
Department of Cell Genetics, Genentech, Inc., South San Francisco, CA 94080.
Henner
D J
DJ
D000595:Amino Acid Sequence; D017104:Bacteriophage M13 / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000235:genetics; D007334:Insulin-Like Growth Factor I / Q000276:immunology*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000276:immunology; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007334:Insulin-Like Growth Factor I; D009838:Oligodeoxyribonucleotides; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins