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Biophysical characterization and single‐chain Fv construction of a neutralizing antibody to measles virus
Author(s) -
Tadokoro Takashi,
Jahan Mst Lubna,
Ito Yuri,
Tahara Maino,
Chen Surui,
Imai Atsutoshi,
Sugimura Natsumi,
Yoshida Koki,
Saito Mizuki,
Ose Toyoyuki,
Hashiguchi Takao,
Takeda Makoto,
Fukuhara Hideo,
Maenaka Katsumi
Publication year - 2020
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.14991
Subject(s) - measles virus , antibody , chemistry , monoclonal antibody , neutralizing antibody , microbiology and biotechnology , epitope , virology , neutralization , hemagglutinin (influenza) , virus , biology , measles , vaccination , genetics
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 n m 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 μ m level. Like 2F4 Fab, scFv inhibited receptor binding and viral entry. This indicates that 2F4 mA b 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.