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

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.
Yu Isoda 1,  Tomohiro Tanaka 1,  Hiroyuki Suzuki 2,  Teizo Asano 1,  Takeo Yoshikawa 3,  Kaishi Kitamura 2,  Yuma Kudo 2,  Ryo Ejima 2,  Kazuki Ozawa 2,  Mika K Kaneko 1,  Yukinari Kato 1 2 3
Affiliations

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.

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