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

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 × 105 independent clones). We first constructed the Vε cDNA library (4.6 × 105 independent clones) and Vκ cDNA library (3.0 × 105 independent clones). Then, the Vε and Vκ 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 Vε from this library was analyzed and proved. Fab clones with high specificity to TCS have been screened out.