Development of a Novel Anti-Mouse CCR2 Monoclonal Antibody (C <sub>2</sub> Mab-6) by N-Terminal Peptide Immunization
Tomohiro Tanaka, Guanjie Li, Teizo Asano, Masaki Saito, Mika K. Kaneko, Hiroyuki Suzuki, Yukinari Kato
Abstract
The CC chemokine receptor type-2 (CCR2) belongs to the G-protein-coupled receptor superfamily, expressed on the cell surface of immune cells and tumors. CCR2 binds to the CC motif chemokine 2/monocyte chemoattractant protein-1, a CC chemokine, which is produced by various cells, including immune-related cells and tumors. Therefore, the development of sensitive monoclonal antibodies (mAbs) for CCR2 has been desired for treatment and diagnosis. This study established a novel, specific, and sensitive anti-mouse CCR2 (mCCR2) mAb; C 2 Mab-6 (rat IgG 1 , kappa), using the mCCR2 synthetic peptide immunization method. C 2 Mab-6 reacted with mCCR2-overexpressed Chinese hamster ovary-K1 cells and L1210 (murine leukemia) cells, which express endogenous mCCR2 in flow cytometry. Furthermore, C 2 Mab-6 showed a high binding affinity for both cells. Hence, C 2 Mab-6 can be a useful tool for analyzing mCCR2-related biological responses, using flow cytometry.