open access

https://hdl.handle.net/2115/78836

Biophysical characterization and single-chain Fv construction of a neutralizing antibody to measles virus

英語

surface plasmon resonance

2019-07-09

John Wiley & Sons

FEBS Journal

287(1)

145-159

The measles virus (MV) is a major cause of childhood morbidity and mortality worldwide. We previously established a mouse monoclonal antibody, 2F4, which shows high neutralizing titers against eight different genotypes of MV. However, the molecular basis for the neutralizing activity of the 2F4 antibody remains incompletely understood. Here, we have evaluated the binding characteristics of a Fab fragment of the 2F4 antibody. Using the MV infectious assay, we demonstrated that 2F4 Fab inhibits viral entry via either of two cellular receptors, SLAM and Nectin4. Surface plasmon resonance (SPR) analysis of recombinant proteins indicated that 2F4 Fab interacts with MV hemagglutinin (MV-H) with a K-D value at the nm level. Furthermore, we designed a single-chain Fv fragment of 2F4 antibody as another potential biopharmaceutical to target measles. The stable 2F4 scFv was successfully prepared by the refolding method and shown to interact with MV-H at the mu m level. Like 2F4 Fab, scFv inhibited receptor binding and viral entry. This indicates that 2F4 mAb uses the receptor-binding site and/or a neighboring region as an epitope with high affinity. These results provide insight into the neutralizing activity and potential therapeutic use of antibody fragments for MV infection.

学術雑誌論文

Accepted Manuscript

This is the peer reviewed version of the following article: T.Tadokoro, et al. / Biophysical characterization and single‐chain Fv construction of a neutralizing antibody to measles virus. FEBS Journal, First published: 09 July 2019, which has been published in final form at https://doi.org/10.1111/febs.14991. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.

1742-464X