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

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: SCIENTIFIC REPORTS; v. 11, n. 1 JAN 14 2021.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 12/10623-2 - Construção e caracterização de scFv anti-FGF2 a partir do hibridoma secretor do Mab 3F12E7
• Beneficiário: Jane Zveiter de Moraes
• Modalidade de apoio: Auxílio à Pesquisa - Regular