Immunoreactivity of humanized single-chain variable fragment against its functional epitope on domain 1 of CD147
Issued Date
2022-12-01
Resource Type
eISSN
20452322
Scopus ID
2-s2.0-85128855102
Pubmed ID
35468972
Journal Title
Scientific Reports
Volume
12
Issue
1
Rights Holder(s)
SCOPUS
Bibliographic Citation
Scientific Reports Vol.12 No.1 (2022)
Suggested Citation
Intasai N., Rangnoi K., Yamabhai M., Pamonsupornwichit T., Thongkum W., Yasamut U., Chupradit K., Takheaw N., Nimmanpipug P., Tayapiwatana C. Immunoreactivity of humanized single-chain variable fragment against its functional epitope on domain 1 of CD147. Scientific Reports Vol.12 No.1 (2022). doi:10.1038/s41598-022-10657-3 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/87546
Title
Immunoreactivity of humanized single-chain variable fragment against its functional epitope on domain 1 of CD147
Author's Affiliation
Other Contributor(s)
Abstract
Domain 1 of CD147 participates in matrix metalloproteinase (MMP) production and is a candidate for targeted therapy to prevent cancer invasion and metastasis. A functional mouse anti-CD147 monoclonal antibody, M6-1B9, was found to recognize domain 1 of CD147, and its respective mouse single-chain variable fragment (ScFvM61B9) was subsequently generated. The EDLGS epitope candidate for M6-1B9 was identified using the phage display peptide technique in this study. For future clinical applications, humanized ScFv specific to domain 1 of CD147 (HuScFvM61B9) was partially adopted from the hypervariable sequences of parental mouse ScFvM61B9 and grafted onto suitable human immunoglobulin frameworks. Molecular modelling and simulation were performed in silico to generate the conformational structure of HuScFvM61B9. These results elucidated the amino acid residues that contributed to the interactions between CDRs and the epitope motif. The expressed HuScFvM61B9 specifically interacted with CD147 at the same epitope as the original mAb, M6-1B9, and retained immunoreactivity against CD147 in SupT1 cells. The reactivity of HuScFvM61B9 was confirmed using CD147 knockout Jurkat cells. In addition, the inhibitory effect of HuScFvM61B9 on OKT3-induced T-cell proliferation as M6-1B9 mAb was preserved. As domain 1 is responsible for cancer invasion and metastasis, HuScFvM61B9 would be a candidate for cancer targeted therapy in the future.