Development of a Novel Anti-Mouse CCR6 Monoclonal Antibody (C6Mab-13) by N-Terminal Peptide Immunization

Monoclon Antib Immunodiagn Immunother. 2022 Dec;41(6):343-349. doi: 10.1089/mab.2022.0021. Epub 2022 Nov 16.

Abstract

The CC chemokine receptor 6 (CCR6) is a G protein-coupled receptor family member that is highly expressed in B lymphocytes, certain subsets of effector and memory T cells, and immature dendritic cells. CCR6 has only one chemokine ligand, CCL20. The CCL20-CCR6 axis has been recognized as a therapeutic target for autoimmune diseases and tumor. This study developed specific monoclonal antibodies (mAbs) against mouse CCR6 (mCCR6) using the peptide immunization method. The established anti-mCCR6 mAb, C6Mab-13 (rat IgG1, kappa), reacted with mCCR6-overexpressed Chinese hamster ovary-K1 (CHO/mCCR6), and mCCR6-endogenously expressed P388 (mouse lymphoid neoplasma) and J774-1 (mouse macrophage-like) cells in flow cytometry. The dissociation constant (KD) of C6Mab-13 for CHO/mCCR6 cells was determined to be 2.8 × 10-9 M, indicating that C6Mab-13 binds to mCCR6 with high affinity. In summary, C6Mab-13 is useful for detecting mCCR6-expressing cells through flow cytometry.

Keywords: monoclonal antibody; mouse CCR6; peptide immunization.

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Immunization
  • Macrophage Inflammatory Proteins* / metabolism
  • Rats
  • Receptors, CCR6* / metabolism

Substances

  • Macrophage Inflammatory Proteins
  • Receptors, CCR6
  • Antibodies, Monoclonal