Epitope Mapping of the Novel Anti-Human CCR9 Monoclonal Antibody (C₉Mab-11) by 2 × Alanine Scanning.

Q3 Medicine

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2023-04-01 DOI:10.1089/mab.2022.0035

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

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引用次数: 1

Abstract

We recently developed a novel anti-human C-C chemokine receptor 9 (hCCR9) monoclonal antibody (mAb), C₉Mab-11, which is applicable to flow cytometry, western blotting, and enzyme-linked immunosorbent assay (ELISA). This study aims to identify the binding epitope of C₉Mab-11 by using 1 × and 2 × alanine (or glycine) substituted-hCCR9 peptides (1 × and 2 × Ala-scan) by ELISA. According to the 1 × Ala-scan analysis, the response of C₉Mab-11 was diminished against M13A of the hCCR9 peptide, but was not eliminated. In the 2 × Ala-scan analysis, the reactions were abolished in the substitution of P11A-N12A, N12A-M13A, and M13A-A14G of hCCR9 N-terminal peptides. The results indicate that the binding epitope of C₉Mab-11 includes Pro11, Asn12, Met13, and Ala14 of hCCR9, with the region around Met13 being particularly important. The successful identification of the C₉Mab-11 epitope might be useful for the future pathophysiological analysis of hCCR9.

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新型抗人CCR9单克隆抗体(C9Mab-11)的2 ×丙氨酸扫描表位定位

我们最近开发了一种新的抗人C-C趋化因子受体9 (hCCR9)单克隆抗体(mAb)， C9Mab-11，适用于流式细胞术，western blotting和酶联免疫吸附测定(ELISA)。本研究的目的是利用酶联免疫吸附法(ELISA)利用1 ×和2 ×丙氨酸(或甘氨酸)取代的hccr9肽(1 ×和2 × Ala-scan)鉴定C9Mab-11的结合表位。根据1 × Ala-scan分析，C9Mab-11对hCCR9肽的M13A的反应减弱，但未完全消除。在2 × Ala-scan分析中，hCCR9 n端肽的P11A-N12A、N12A-M13A和M13A-A14G被取代，反应被取消。结果表明，C9Mab-11的结合表位包括hCCR9的Pro11、Asn12、Met13和Ala14，其中Met13周围的区域尤为重要。C9Mab-11表位的成功鉴定可能为今后hCCR9的病理生理分析提供依据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Medicine-Immunology and Allergy

CiteScore

4.80

自引率

0.00%

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