Cx₄Mab-1: a Novel Anti-mouse CXCR4 Monoclonal Antibody for Flow Cytometry

Abstract

The CXC chemokine receptor 4 (CXCR4, CD184) is a member of the G protein-coupled receptor family that is expressed in most leukocytes. Overexpression of CXCR4 is associated with poor prognosis in not only hematopoietic malignancy but also solid tumors. Because CXCR4 is an attractive target for tumor therapy, reliable preclinical murine models using anti-CXCR4 monoclonal antibodies (mAbs) have been warranted. This study established a novel anti-mouse CXCR4 (mCXCR4) mAb using the Cell-Based Immunization and Screening (CBIS) method. Flow cytometric analysis showed that an anti-mCXCR4 mAb, Cx4Mab-1 (rat IgG2a, kappa), recognized mCXCR4-overexpressed Chinese hamster ovary-K1 (CHO/mCXCR4) cells and endogenously mCXCR4-expressing mouse myeloma P3X63Ag8U.1 (P3U1) cells. Furthermore, Cx4Mab-1 did not recognize mCXCR4-knockout P3U1 cells. The dissociation constants of Cx4Mab-1 for CHO/mCXCR4 and P3U1 were determined as 6.4 × 10−9 M and 2.3 × 10-9 M, respectively, indicating that Cx4Mab-1 possesses a high affinity to both endogenous and exogenous mCXCR4-expressing cells. These results indicate that Cx4Mab-1 could be a useful tool for preclinical mouse models.

Cx4Mab-1: A Novel Anti-Mouse CXCR4 Monoclonal Antibody for Flow Cytometry

“…Next, we have been trying to establish the anti-C-X-C motif chemokine receptor mAbs. We recently obtained an anti-mouse CXCR3 mAb [29], an anti-mouse CXCR4 mAb [30], an anti-mouse CXCR6 mAb [31]. In this study, we have successfully developed anti-mouse CXCR1 (mCXCR1) mAbs using the CBIS method which is applicable to flow cytometry and western blot analysis.…”