Generation and functional characterization of a single-chain variable fragment (scFv) of the anti-FGF2 3F12E7 monoclonal antibody
| Autor: | Katia Regina Perez, Bruno Andrade Costa, Jane Zveiter de Moraes, Rodrigo Barbosa de Aguiar, Vitor Oliveira, Tábata de Almeida da Silva, Camila Braggion, Renata Yoshiko Yamada, Marcelo A. Mori, Marcelo F.M. Machado |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
medicine.drug_class Science In silico Angiogenesis Inhibitors Immunoglobulin light chain Monoclonal antibody Article Immunoglobulin G law.invention 03 medical and health sciences Antineoplastic Agents Immunological 0302 clinical medicine law medicine Humans Single-chain variable fragment Multidisciplinary biology Chemistry Proteins respiratory system Neoplasm Proteins 030104 developmental biology Biochemistry 030220 oncology & carcinogenesis biology.protein Recombinant DNA Medicine Fibroblast Growth Factor 2 Immunotherapy Antibody Linker Single-Chain Antibodies |
| Zdroj: | Scientific Reports, Vol 11, Iss 1, Pp 1-11 (2021) Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-020-80746-8 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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