The preparation and tumor cell apoptosis-inducing activity assay of anti-human DR5 single-chain antibody

Purpose To construct,express,purify anti-human DR5 scFv,and to assay its activity of apoptosis induction for tumor cell lines.MethodsVariable region sequences of heavy chain and light chain to murine anti-human DR5 monoclonal antibody were acquired by RT-PCR,then they were linked through a flexible linker peptide and was cloned into the expression vector.After being expressed by E.coli and purified using affinity chromatography,the recombinant protein was employed to MTT assay as well as apoptosis assay kit in order to detect its apoptosis-inducing activity.ResultsThe sequences we′ve got were established as the variable region genes of antibody′s heavy and light chain,while the recombinant proteins expressed and purified possessing the apoptosis-inducing activity come near to complete antibody.ConclusionAnti-human DR5 scFv can supply material for tumor immunology research as drug candidate inducing tumor cell apoptosis.