Development of an Anti-human CCR2 Monoclonal Antibody (C2Mab-9) by N-Terminal Peptide Immunization.

Abstract

The CC chemokine receptor type-2 (CCR2) is one of the members of the G protein-coupled receptor superfamily, which are expressed on the cell surface of immune and tumor cells. CCR2 binds to the C-C motif chemokine ligand 2 (CCL2)/monocyte chemoattractant protein-1 (MCP-1), which is produced by various cells, including tumor and immune-related cells. Therefore, the development of sensitive monoclonal antibodies (mAbs) for CCR2 has been desired for treatment and diagnosis. In this study, we established a specific antihuman CCR2 (hCCR2) mAb, C₂Mab-9 (mouse IgG₁, kappa), using the synthetic peptide immunization method. Flow cytometric and immunocytochemical results showed that C₂Mab-9 reacted with hCCR2-expressing U937 (human histiocytic lymphoma) and natural killer cells. Furthermore, C₂Mab-9 showed the moderate binding affinity for both cells. Conclusively, C₂Mab-9 can be a useful tool for analyzing hCCR2-related biological responses.