The construction and expression of phage display library of anti human lung adenocarcinoma monoclonal antibody 5F-11

Objective: To construct and express a phage display library of anti human lung cancer monoclonal antibody 5F-11. Methods: Immunoglobulin variable regions (VH, VL) were amplified from 5F-11 hybridrom by RT-PCR. ScFv genes consisting of VH DNA and VL DNA joined together by a linker DNA were cloned into a phage vector pCANT-AB5E. After 4 rounds of screening with lung adenocarcinoma cell line A2 as antigen, an enriched secondary phage display library was obtained. Results: A recombinant phage display library with total of 8 × 107 pfu/ml was established. Randomized clones from unselected library digested with BstN I showed different patterns, however, those from selected library showed that phages with special pattern were enriched. Twenty-three out of 30 clones were found to respond strongly to A2 cell lines. Conclusion: The ScFv of anti-lung adenocarcinoma monoclonal antibody 5F-11 can be successfully produced, which may be useful to widen the application of the antibody.