Isolation and characterization of full-length genes encoding the anti-human CD45 antibody from the hybridoma cell line 16E8-F2

Abstract

Though the hybridoma technology has been widely applied in the production of monoclonal antibodies, it has existed some disadvantages including low yield and genetic instability. Therefore, an alternative approach should be taken into account. Recently, recombinant monoclonal antibody technology has emerged as the best choice to cure hybridoma related drawbacks. However, recombinant antibodies require known genes for their generation. The purpose of this study is to collect the full-length genes encoding the anti-human CD45 antibody derived from the hybridoma cell line 16E8-F2. In this research, we designed specific primer pairs to amplify the light and heavy chain genes of the antibody through the PCR method. Afterward, the genes were separately cloned into a cloning vector called pJET1.2/blunt. The generated recombinant pJET1.2 vectors will serve as the main material source to manufacture the recombinant monoclonal antibody recognizing human CD45 protein tomorrow.