Fine epitope mapping of a human disulfide-stabilized diabody against fibroblast growth factor-2

• Published in: Journal of biochemistry (Tokyo)
• Main Authors: Zhong, Jiangchuan, Zhang, Simin, Zhang, Ligang, Cai, Yaxiong, Deng, Yanrui, Zheng, Qubo, Deng, Ning
• Format: Journal Article
• Language: English
• Published: England 31.12.2018
• ISSN: 1756-2651

The human fibroblast growth factor-2 (FGF-2) highly expressed in tumors is an important factor to promote tumor angiogenesis and lymphangiogensis. A disulfide-stabilized diabody (ds-Diabody) could specifically target FGF-2 and show its advantages in inhibition of tumor angiogenesis and growth. It is very important for antibody drugs to confirm the fine epitope. Here, theoretical structure models of FGF-2 and antibody were built by homology modeling. The amino acid residues in the interaction interface of antigen and antibody were analyzed by molecular docking. The potential epitope was predicted by homology modeling and molecular docking of antigen-antibody and site-directed mutation assays of alanine scanning. The predicted epitope was verified by antigen mutagenesis and enzyme-linked immunosorbent assay (ELISA). The epitope mapping assay showed that the epitope of ds-Diabody against FGF-2 was defined by the discontinuous sites including six amino acid residues (P23, Q65, R69, G70, Y82 and R118). The results showed that the epitope was localized in the interaction interface of FGF-2 and ds-Diabody. The fine epitope mapping provided the important information for understanding the inhibition activity of ds-Diabody against FGF-2 and helping in the further development of ds-Diabody against FGF-2 as a potentially promising antibody drug for future cancer therapy.