Expression of Anti-RBC Blood Group B Substance ScFv by Using Phage Display Technology

Antibody library technology coupled with guide selection is one of the efficient methods to generate antigen-specific antibodies. In this paper, the recombinant phage display technique was used to construct, clone screen, and express single chain antibody of 5D12 hybridoma. The variable region genes of antibody were amplified by PCR from a hybridoma cell line 5D12 which secreted group B substance. The ScFv gene fragments were successfully cloned into phagemid vector pHEN-1. Rescuing with M13KO7 helper phage, a recombinant phage ScFv library with titer of 3 × 109 pfu/ml was established. Panned by whole blood group B cell over three rounds, the positive recombinant phages were sequenced and identified by immunol-blot assay. The result of immunoblot indicated that the phage-displayed 5D12-ScFv antibody retained the affinity and specificity of the original intact antibody to blood group B substance, which would be potentially useful in constructing engineering antibody fragments as blood grouping reagent.