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.
Highlights
Single-chain variable fragments are small-sized artificial constructs composed of the immunoglobulin heavy and light chain variable regions connected by a peptide linker
ScFvs consist of artificial constructs composed of the variable regions of the heavy ( VH) and light (VL) immunoglobulin chains connected by a flexible peptide linker in a way to retain the antigen-binding properties of the corresponding immunoglobulin G (IgG) antibody[2]
Our molecular construct follows a traditional scFv scheme comprising the VH and VL segments of the IgG counterpart. These domains were joined by a 15-residue glycine-serine linker, used in many of the reported scFvs[13, 15,16,17,18], leading to an artificial VH-VL arrangement – named 3F12E7 scFv – whose monomer molecular mass is around 35 kDa
Summary
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. Abbreviations CDR Complementarity-determining region ctrl Control FGF2 Fibroblast growth factor 2 HUVEC Human umbilical vein endothelial cell IgG Immunoglobulin G mAb Monoclonal antibody scFv Single-chain variable fragment VH Variable heavy domain VL Variable light domain
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
