Construction and Screening of a Phage Display Library of Repertoire Single Chain Antibody from Mouse Immunized with vWF-A1

To screen the monoclonal antibody with high affinity to vWF-Al fragment, phage display library was constructed from mouse immunized with this fragment to lay a foundation for its clinical usage. The heavy chain and light chain variable region (VH and NL ) gene repertoire of immunoglobulin were amplified from mRNA of spleen cells by RT-PCR, and joined by a DNA linker encoding peptide as a single chain Fv (ScFv )fragment with overlap extension. These fragments were cloned into phage vector pHENl and the phage display library was constructed. The affinity selection and ELISA assay were adopted for the identification of specific phage antibody to vWF-Al. Results showed that the high affinity ScFv was obtained after 5 rounds of panning, with its DNA sequence conforming to that of mouse antibody. It concludes that the phage display library of repertoire single chain anti-vWF-Al antibody was successfully constructed and the single chain antibody with high affinity to vWF-Al was screened.