Clone of the anti-DR5 antibody variable sequence by phage surface display

Objective:To obtain the gene sequence of antibody against human death receptor 5 by the phage surface display.Methods:By the technique of phage display,the heavy chain and light chain variable region genes(VH and VL) were assembled into single chain antibodies(scFv) with overlap extension reaction by L inker primer mix.The scFv were cloned into PAK100 phagemid and introduced into E.coli XL1-Blue.The phagemid containing bacterial colonies were infected with VSCM13,then the total surface display library of anti-human-death receptor 5 was established.The library was screened by death receptor 5 to get the antigen-specific phage clones.Positive clones were characterized by DNA sequencing.Results:After three panning rounds ofbinding-elution-amplification,10 individual clones were randomly selected and identified by enzyme-immunosorbent assay(ELISA),4 of which has the characteristic of DR5.Conclusion:The technique of phage display is a more creditable method to obtain antibody gene sequence.