Protein III-based single-chain antibody phage display using bacterial cells bearing an additional genome of a gene-III-lacking helper phage

Biotechniques. 2007 Jun;42(6):760-5. doi: 10.2144/000112461.

Abstract

We present herein a novel method of pIII-based antibody phage display using Hpd3cells--bacterial cells bearing the genome of a gene-III-lacking helper phage (VCSM13d3). A high level of single-chain variable fragments (scFvs) was displayed in the consequent phagemid particles using Hpd3cells to rescue the phagemid encoding scFv-pIII. Hpd3cells considerably improved the specific enrichment factor when used for constructing an immunized antibody library. In addition, using Hpd3cells could overcome pill resistance and can contribute to the efficient enrichment of specific binding antibodies from a phage display library, thereby increasing the chance of obtaining more diverse antibodies specific for target antigens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / genetics*
  • Antibodies / immunology
  • Bacteriophage M13 / immunology*
  • Escherichia coli / genetics
  • Genome, Bacterial*
  • Immunoglobulin Fragments / genetics
  • Immunoglobulin Variable Region / genetics*
  • Models, Genetic
  • Peptide Library*
  • Plasmids

Substances

  • Antibodies
  • Immunoglobulin Fragments
  • Immunoglobulin Variable Region
  • Peptide Library