Selection of recombinant antibodies from antibody gene libraries

Methods Mol Biol. 2014;1101:305-20. doi: 10.1007/978-1-62703-721-1_14.

Abstract

Antibodies are indispensable detection reagents for research and diagnostics and represent the biggest class of biological therapeutics on the market. In vitro antibody selection systems offer many advantages over animal-based technologies because the whole selection process is independent of the in vivo immune response. In the last two decades antibody phage display has evolved to the most robust and widely used method and has already yielded thousands of antibodies. The selection of binders by phage display is also referred to as "panning" and based on the specific molecular interaction of antibody phage with an immobilized antigen thus allowing the enrichment and isolation of antigen-specific monoclonal binders from very large antibody gene libraries. Here, we give detailed protocols for the selection of recombinant antibody fragments from antibody gene libraries in microtiter plates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal, Murine-Derived / genetics
  • Antibodies, Monoclonal, Murine-Derived / immunology
  • Antibodies, Monoclonal, Murine-Derived / isolation & purification*
  • Antibody Specificity
  • Antigens / immunology
  • Cell Surface Display Techniques
  • Enzyme-Linked Immunosorbent Assay
  • Gene Library*
  • Humans
  • Immobilized Proteins / immunology
  • Mice
  • Protein Binding
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification

Substances

  • Antibodies, Monoclonal, Murine-Derived
  • Antigens
  • Immobilized Proteins
  • Recombinant Proteins