Selection of an anti-IGF-1 Fab from a Fab phage library created by mutagenesis of multiple CDR loops

Gene. 1993 Jun 15;128(1):103-9. doi: 10.1016/0378-1119(93)90160-5.

Abstract

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.

MeSH terms

  • Amino Acid Sequence
  • Bacteriophage M13 / genetics*
  • Base Sequence
  • Cloning, Molecular
  • Humans
  • Immunoglobulin Fab Fragments / biosynthesis*
  • Immunoglobulin Fab Fragments / genetics
  • Immunoglobulin Heavy Chains / biosynthesis
  • Immunoglobulin Heavy Chains / genetics
  • Insulin-Like Growth Factor I / immunology*
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides
  • Plasmids
  • Protein-Tyrosine Kinases / genetics
  • Proto-Oncogene Proteins / immunology
  • Receptor, ErbB-2
  • Recombinant Fusion Proteins / biosynthesis

Substances

  • Immunoglobulin Fab Fragments
  • Immunoglobulin Heavy Chains
  • Oligodeoxyribonucleotides
  • Proto-Oncogene Proteins
  • Recombinant Fusion Proteins
  • Insulin-Like Growth Factor I
  • Protein-Tyrosine Kinases
  • Receptor, ErbB-2