Construction of Human Antibody Gene Libraries and Selection of Antibodies by Phage Display

Methods Mol Biol. 2014;1060:215-43. doi: 10.1007/978-1-62703-586-6_12.


Antibody phage display is the most commonly used in vitro selection technology and has yielded thousands of useful antibodies for research, diagnostics, and therapy.The prerequisite for successful generation and development of human recombinant antibodies using phage display is the construction of a high-quality antibody gene library. Here, we describe the methods for the construction of human immune and naive scFv gene libraries.The success also depends on the panning strategy for the selection of binders from these libraries. In this article, we describe a panning strategy that is high-throughput compatible and allows parallel selection in microtiter plates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal, Humanized / genetics*
  • Antibodies, Monoclonal, Humanized / immunology*
  • Antibody Specificity / immunology
  • Cell Surface Display Techniques*
  • Gene Library*
  • Humans
  • Peptide Library*
  • Protein Engineering / methods
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology


  • Antibodies, Monoclonal, Humanized
  • Peptide Library
  • Single-Chain Antibodies