Production of Genetically Engineered Antigen Specific Human Monoclonal IgE Antibody

Abstract

We established B cell clones, AC7–1/F9, CB7–1/E2 and CB7–8/F5 producing rice allergenic protein (RA)-specific human monoclonal IgM antibodies. In this study, we attempted to generate antigen specific human IgE antibody by genetic engineering of IgM antibody created by in vitro immunization. The single chain variable fragments (ScFv). were generated by PCR techniques. The DNA fragments of antigen specific were rescued 63 of recombinant phage clones after selection of ScFv for antigen specificity to RA by phage siaplay method. The antigen specific VH and constnat region of human IgE (Cs) DNAs were fused in frame, and inserted into the downstream of CMV promoter of mamalian expression vector. The antigen specific Vλ or Vk fused to corresponding constant region were independently inserted into the downstream of CMV promoter of the extension vector, pIRESbleo. These constructs were simultaneously transfected into CHO cell line, and expression of IgE constant region mRNA and specificity for RA of IgE producing cells were determined.