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

Takashi Tadokoro, Mst Lubna Jahan, Yuri Ito, Maino Tahara, Surui Chen, Atsutoshi Imai, Natsumi Sugimura, Koki Yoshida, Mizuki Saito, Toyoyuki Ose, Takao Hashiguchi, Makoto Takeda, Hideo Fukuhara, Katsumi Maneaka

研究成果: ジャーナルへの寄稿記事

抄録

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 KD 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 μm 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.

元の言語英語
ジャーナルFEBS Journal
DOI
出版物ステータス受理済み/印刷中 - 1 1 2019

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Single-Chain Antibodies
Measles virus
Neutralizing Antibodies
Viruses
Antibodies
Immunoglobulin Variable Region
Immunoglobulin Fragments
Immunoglobulin Fab Fragments
Surface Plasmon Resonance
Measles
Hemagglutinins
Surface plasmon resonance
Therapeutic Uses
Virus Diseases
Recombinant Proteins
Epitopes
Assays
Binding Sites
Genotype
Monoclonal Antibodies

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Tadokoro, T., Jahan, M. L., Ito, Y., Tahara, M., Chen, S., Imai, A., ... Maneaka, K. (受理済み/印刷中). Biophysical characterization and single-chain Fv construction of a neutralizing antibody to measles virus. FEBS Journal. https://doi.org/10.1111/febs.14991

Biophysical characterization and single-chain Fv construction of a neutralizing antibody to measles virus. / 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; Maneaka, Katsumi.

:: FEBS Journal, 01.01.2019.

研究成果: ジャーナルへの寄稿記事

Tadokoro, T, Jahan, ML, Ito, Y, Tahara, M, Chen, S, Imai, A, Sugimura, N, Yoshida, K, Saito, M, Ose, T, Hashiguchi, T, Takeda, M, Fukuhara, H & Maneaka, K 2019, 'Biophysical characterization and single-chain Fv construction of a neutralizing antibody to measles virus', FEBS Journal. https://doi.org/10.1111/febs.14991
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 ; Maneaka, Katsumi. / Biophysical characterization and single-chain Fv construction of a neutralizing antibody to measles virus. :: FEBS Journal. 2019.
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AU - Chen, Surui

AU - Imai, Atsutoshi

AU - Sugimura, Natsumi

AU - Yoshida, Koki

AU - Saito, Mizuki

AU - Ose, Toyoyuki

AU - Hashiguchi, Takao

AU - Takeda, Makoto

AU - Fukuhara, Hideo

AU - Maneaka, Katsumi

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