Generation and functional characterization of a single-chain variable fragment (scFv) of the anti-FGF2 3F12E7 monoclonal antibody

Author(s)

de Aguiar, Rodrigo Barbosa ^[1] ; da Silva, Tabata de Almeida ^[1] ; Costa, Bruno Andrade ^[1] ; Marcondes Machado, Marcelo Ferreira ^[1] ; Yamada, Renata Yoshiko ^[1] ; Braggion, Camila ^[1] ; Perez, Katia Regina ^[1] ; Silva Mori, Marcelo Alves ^[2] ; Oliveira, Vitor ^[1] ; de Moraes, Jane Zveiter ^[1]

Affiliation

^[1] Univ Fed Sao Paulo, Dept Biophys, Escola Paulista Med, Rua Tres Maio, 100 Vila Clementino, BR-04044020 Sao Paulo, SP - Brazil

^[2] Univ Estadual Campinas, Inst Biol, Campinas - Brazil

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 11, n. 1 JAN 14 2021.
• Web of Science Citations: 0

Abstract

Single-chain variable fragments (scFvs) are small-sized artificial constructs composed of the immunoglobulin heavy and light chain variable regions connected by a peptide linker. We have previously described an anti-fibroblast growth factor 2 (FGF2) immunoglobulin G (IgG) monoclonal antibody (mAb), named 3F12E7, with notable antitumor potential revealed by preclinical assays. FGF2 is a known angiogenesis-associated molecule implicated in tumor progression. In this report, we describe a recombinant scFv format for the 3F12E7 mAb. The results demonstrate that the generated 3F12E7 scFv, although prone to aggregation, comprises an active anti-FGF2 product that contains monomers and small oligomers. Functionally, the 3F12E7 scFv preparations specifically recognize FGF2 and inhibit tumor growth similar to the corresponding full-length IgG counterpart in an experimental model. In silico molecular analysis provided insights into the aggregation propensity and the antigen-recognition by scFv units. Antigen-binding determinants were predicted outside the most aggregation-prone hotspots. Overall, our experimental and prediction dataset describes an scFv scaffold for the 3F12E7 mAb and also provides insights to further engineer non-aggregated anti-FGF2 scFv-based tools for therapeutic and research purposes. (AU)

• FAPESP's process: 12/10623-2 - Construction and characterization of anti-FGF-2 scFv from 3F12E7 hybridoma
• Grantee: Jane Zveiter de Moraes
• Support Opportunities: Regular Research Grants