Construction of human phage antibody library and screening of phage antibodies against bFGF

Objective To construct human phage antibody library from autoimmunity patients' blood containing high titer of anti-bFGF antibody and select anti-bFGF phage antibody from the library. Methods The heavy chain and light chain of the Fab fragments of human IgG were separately amplified by RT-PCR from peripheral lymphocytes of autoimmunity patients' blood. The amplified Fd chain and light chain were inserted into vector pComb3 and electro-transferred into E.coli XL1-Blue cells. The recombinant phage was rescued by coculture with helper phage VCSM13 for constructing human phage antibody library. After three rounds of panning, the specific phage antibodies against bFGF were highly enriched. The high affinity phage antibodies against bFGF were identified by indirect ELISA. Results The results shown that the human phage antibody library was constructed (library content was 2.5×10~7) and specific phage antibodies were obtained from the antibody library. Conclusion The construction of the human phage antibody library and the preparation of the anti-bFGF phage antibodies provide a basis for the research of anti-bFGF drugs.