Guided selection methods through chain shuffling

Methods Mol Biol. 2009:562:133-42. doi: 10.1007/978-1-60327-302-2_10.

Abstract

We provide procedures for the panning of fully humanized Fab antibodies using guided selection. Human heavy and light chain genes are amplified. A parental light chain is cloned into a phage display vector and combined with the heavy chain library. After several rounds of panning, positive clones are identified and the heavy chain sequences that are recovered are combined with light chains for further selection by phage display. Human Fab antibodies are obtained that bind the same epitope as the parental antibody.

MeSH terms

  • Humans
  • Immunoglobulin Fab Fragments / genetics*
  • Immunoglobulin Fab Fragments / immunology
  • Immunoglobulin Fab Fragments / isolation & purification*
  • Immunoglobulin Heavy Chains / genetics*
  • Immunoglobulin Heavy Chains / immunology
  • Immunoglobulin Light Chains / genetics*
  • Immunoglobulin Light Chains / immunology
  • Peptide Library*
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / immunology

Substances

  • Immunoglobulin Fab Fragments
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Light Chains
  • Peptide Library
  • Recombinant Fusion Proteins