TY - JOUR
T1 - Epitope Mapping of an Anti-Mouse CCR2 Monoclonal Antibody (C2Mab-6) Using Enzyme-Linked Immunosorbent Assay
AU - Tanaka, Tomohiro
AU - Suzuki, Hiroyuki
AU - Asano, Teizo
AU - Li, Guanjie
AU - Nanamiya, Ren
AU - Tateyama, Nami
AU - Isoda, Yu
AU - Okada, Yuki
AU - Kobayashi, Hiyori
AU - Yoshikawa, Takeo
AU - Kaneko, Mika K.
AU - Kato, Yukinari
N1 - Funding Information:
This research was supported in part by Japan Agency for Medical Research and Development (AMED) under Grant Nos. JP22ama121008 (to Y.K.), JP22am0401013 (to Y.K.), JP22bm1004001 (to Y.K.), JP22ck0106730 (to Y.K.), and JP21am0101078 (to Y.K.).
Publisher Copyright:
© Copyright 2022, Mary Ann Liebert, Inc., publishers 2022.
PY - 2022/12/1
Y1 - 2022/12/1
N2 - CC chemokine receptor type-2 (CCR2) is a member of the G protein-coupled receptors, and is mainly expressed on cell surface of immune cells. CCR2 binds to its ligand, C-C motif chemokine 2 (also named as monocyte chemoattractant protein-1), which involves in the tumor progression by modulating the tumor microenvironment. Therefore, the monoclonal antibody (mAb) targeting CCR2 could be one of the strategies for cancer treatment. In this study, we investigated the critical epitope of C2Mab-6, an anti-mouse CCR2 (mCCR2) mAb developed by N-Terminal peptides immunization. We first performed enzyme-linked immunosorbent assay (ELISA) using N-Terminal peptides of mCCR2 and demonstrated that C2Mab-6 recognizes 1-19 amino acids of mCCR2. We further performed ELISA using 20 alanine-substituted peptides of mCCR2. C2Mab-6 lost the reaction to the alanine-substituted peptides of D3A, N4A, M6A, P8A, Q9A, and F10A. These results indicate that the binding epitope of C2Mab-6 includes Asp3, Asn4, Met6, Pro8, Gln9, and Phe10 of mCCR2.
AB - CC chemokine receptor type-2 (CCR2) is a member of the G protein-coupled receptors, and is mainly expressed on cell surface of immune cells. CCR2 binds to its ligand, C-C motif chemokine 2 (also named as monocyte chemoattractant protein-1), which involves in the tumor progression by modulating the tumor microenvironment. Therefore, the monoclonal antibody (mAb) targeting CCR2 could be one of the strategies for cancer treatment. In this study, we investigated the critical epitope of C2Mab-6, an anti-mouse CCR2 (mCCR2) mAb developed by N-Terminal peptides immunization. We first performed enzyme-linked immunosorbent assay (ELISA) using N-Terminal peptides of mCCR2 and demonstrated that C2Mab-6 recognizes 1-19 amino acids of mCCR2. We further performed ELISA using 20 alanine-substituted peptides of mCCR2. C2Mab-6 lost the reaction to the alanine-substituted peptides of D3A, N4A, M6A, P8A, Q9A, and F10A. These results indicate that the binding epitope of C2Mab-6 includes Asp3, Asn4, Met6, Pro8, Gln9, and Phe10 of mCCR2.
KW - CMab-6
KW - enzyme-linked immunosorbent assay
KW - epitope
KW - monoclonal antibody
KW - mouse CCR2
UR - http://www.scopus.com/inward/record.url?scp=85145022653&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85145022653&partnerID=8YFLogxK
U2 - 10.1089/mab.2022.0020
DO - 10.1089/mab.2022.0020
M3 - Article
C2 - 36346278
AN - SCOPUS:85145022653
SN - 2167-9436
VL - 41
SP - 339
EP - 342
JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
IS - 6
ER -
