J Cancer 2018; 9(11):2003-2011. doi:10.7150/jca.24255
Construction of a human monoclonal antibody against bFGF for suppression of NSCLC
1. Guangdong Province Engineering Research Center for antibody drug and immunoassay, College of Life Science and Technology, Jinan University, Guangzhou 510632, Guangdong Province, China.
2. Department of Oncology, the First Affiliated Hospital of Jinan University, Guangzhou 510632, Guangdong Province, China.
3. Akeso Biopharma, Inc., Zhongshan, 528400, Guangdong Province, China
* These authors contributed equally to this work.
Wang S, Qin Y, Wang Z, Xiang J, Zhang Y, Xu M, Li B, Xia Y, Zhang P, Wang H. Construction of a human monoclonal antibody against bFGF for suppression of NSCLC. J Cancer 2018; 9(11):2003-2011. doi:10.7150/jca.24255. Available from http://www.jcancer.org/v09p2003.htm
Compelling evidence implicates that overexpression of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor 1 (FGFR1) in non-small cell lung cancer (NSCLC) drives tumor progression, can serve as prognostic biomarkers or therapeutic targets for NSCLC patients. But at present, we still lack of effective drugs for bFGF. The preparation of monoclonal antibodies against bFGF or to understand its mechanism of action is urgently need. Previously, we used hybridoma technology to produce a murine anti-bFGF monoclonal antibody (E12). However, E12 carries risks of heterogeneity and immunogenicity. In the present work, we produced three humanized variants (H1L1, H2L2 and H3L3) based on E12 by substituting residues in or near the complementarity-determining region (CDR). In addition, we thoroughly explored VH/VL domain combinations to simulate full-length IgG1 antibodies using computational protein design. H3L3 was selected for further study, as it demonstrated the best humanization and strongest affinity for bFGF. Specially, humanization of H3L3's light chain and heavy chain were 100% and 98.89%, respectively. The FGF2 neutralizing effect of H3L3 were confirmed by ELISA. We also found that H3L3 can effectively suppress the growth and angiogenesis of cancer through reduce the phosphorylation of AKT and MAPK. Moreover, H3L3 dramatically reduced tumor size and micro-vessel density in nude mice. Altogether, our study demonstrates that H3L3 exerts anti-tumor effects by impeding NSCLC development.
Keywords: bFGF, Antibody humanization, Non-Small Cell Lung, Targeted therapies