Construction of Fab Fragments Phage Display Library for Human B-lymphoma

Objective To construct Fab phage display library against human B-lymphoma.Me-(thods)The total RNA was extracted from the spleen cells of the immunized BALB/c mice with Raji B-lymphoma cell strain and the immunoglobulin genes of the light chain κ and the heavy chain Fd fragments were amplified by RT-PCR.After restrictive digestion,purification and ligation,the light chain κ and the heavy chain Fd were subsequently inserted into the phagmid vector pComb3H-SS,and then electroprated into E.coli XL1-Blue.Results The Fab phage display library against B-lymphoma was constructed.The combination rate of the light chains and heavy chain Fd fragments was 100% and 78% respectively.The volume of Fab phage library was reached 2.18×10~7.Conclusion The Fab phage display library against human B-lymphoma is constructed successfully.The present study can be used as foundation for succeeding screening of specific antibody against human B-lymphoma.