Epitope Mapping Using the Cell-Based 2 × Alanine Substitution Method About the Anti-mouse CXCR6 Monoclonal Antibody, Cx6Mab-1

Yu Isoda; Tomohiro Tanaka; Hiroyuki Suzuki; Teizo Asano; Takeo Yoshikawa; Kaishi Kitamura; Yuma Kudo; Ryo Ejima; Kazuki Ozawa; Mika K Kaneko; Yukinari Kato

doi:10.1089/mab.2022.0029

Epitope Mapping Using the Cell-Based 2 × Alanine Substitution Method About the Anti-mouse CXCR6 Monoclonal Antibody, Cx₆Mab-1

Monoclon Antib Immunodiagn Immunother. 2023 Feb;42(1):22-26. doi: 10.1089/mab.2022.0029. Epub 2022 Nov 16.

Authors

Yu Isoda¹, Tomohiro Tanaka¹, Hiroyuki Suzuki², Teizo Asano¹, Takeo Yoshikawa³, Kaishi Kitamura², Yuma Kudo², Ryo Ejima², Kazuki Ozawa², Mika K Kaneko¹, Yukinari Kato^{1

2

3}

Affiliations

¹ Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
² Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
³ Department of Pharmacology, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.

PMID: 36383116
DOI: 10.1089/mab.2022.0029

Abstract

An anti-mouse CXC chemokine receptor 6 (mCXCR6) monoclonal antibody (mAb), Cx₆Mab-1, was developed recently. Cx₆Mab-1 is applicable for flow cytometry, Western blotting, and enzyme-linked immunosorbent assay. The purpose of this study is to determine the binding epitope of Cx₆Mab-1 using 2 × alanine mutated mCXCR6. Analysis of flow cytometry revealed that Cx₆Mab-1 did not recognize S8A-A9G, L10A-Y11A, D12A-G13A, and H14A-Y15A mutants of mCXCR6. The results clearly indicate that the binding epitope of Cx₆Mab-1 includes Ser8, Ala9, Leu10, Tyr11, Asp12, Gly13, His14, and Tyr15 of mCXCR6. The successful determination of the Cx₆Mab-1 epitope might contribute to the pathophysiological investigation of mCXCR6.

Keywords: 2 × Ala scanning; epitope; flow cytometry; monoclonal antibody; mouse CXCR6.

MeSH terms

Alanine* / genetics
Antibodies, Monoclonal*
Enzyme-Linked Immunosorbent Assay
Epitope Mapping / methods
Epitopes
Immunosuppressive Agents

Substances

Antibodies, Monoclonal
Alanine
Epitopes
Immunosuppressive Agents