Construction and diversity analysis of a murine IgE phage surface display library

Cell Res. 1997 Dec;7(2):161-70. doi: 10.1038/cr.1997.17.

Abstract

To make further investigation of the IgE antibody repertoire in Trichosanthin (TCS) allergic responses, a murine IgE phage surface display library was constructed (3.0 x 10(5) independent clones). We first constructed the V epsilon cDNA library (4.6 x 10(5) independent clones) and V kappa cDNA library (3.0 x 10(5) independent clones). Then, the V epsilon and V kappa gene segments were amplified from both libraries by PCR respectively, and assembled into Fab fragment by SOE PCR. The phage library containing Fabs was thus constructed. The diversity of V epsilon from this library was analyzed and proved. Fab clones with high specificity to TCS have been screened out.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibody Diversity / genetics*
  • Bacteriophage M13 / genetics*
  • Bacteriophage M13 / immunology*
  • DNA, Complementary / analysis
  • Gene Library*
  • Genetic Vectors
  • Immunoglobulin E / genetics*
  • Immunoglobulin Variable Region / genetics
  • Immunoglobulin epsilon-Chains / genetics
  • Immunoglobulin kappa-Chains / genetics
  • Mice
  • Mice, Inbred C57BL
  • Polymerase Chain Reaction
  • Trichosanthin / immunology

Substances

  • DNA, Complementary
  • Immunoglobulin Variable Region
  • Immunoglobulin epsilon-Chains
  • Immunoglobulin kappa-Chains
  • Immunoglobulin E
  • Trichosanthin