Characterization of a novel adhesion function blocking monoclonal antibody to rat/mouse P-selectin generated in the P-selectin-deficient mouse

Hybridoma. 1997 Jun;16(3):249-57. doi: 10.1089/hyb.1997.16.249.


P-selectin is an important adhesion molecule involved in leukocyte migration. However, to date, no monoclonal antibodies (MAb) generated against rat P-selectin have been identified which block P-selectin mediated leukocyte adhesion. Most studies in the rat have utilized crossreacting antibodies generated against P-selectin in higher species. In a P-selectin deficient mouse we generated an anti-rat/mouse P-selectin MAb, designated RMP-1, by immunization with activated rat platelets. This IgG2a MAb immunoprecipitates a 140 kDa protein under reducing conditions from rat platelet lysate. By ELISA and immunofluorescence flow cytometry, MAb RMP-1 reacts with thrombin-activated but not unactivated rat platelets. In addition, by ELISA MAb RMP-1 binds to activated mouse platelets and recombinant rat and mouse P-selectin. MAb RMP-1 inhibited adhesion of HL-60 myeloid cells to immobilized mouse P-selectin by 97% and to activated rat and mouse platelets by 100% under static conditions, confirming the adhesion function blocking activity of MAb RMP-1. This novel MAb should be useful for studying P-selectin function in vitro and in vivo in both rat and mouse inflammation models.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Blocking / chemistry
  • Antibodies, Blocking / immunology*
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / immunology*
  • Blood Platelets / immunology
  • Blotting, Western
  • Cell Adhesion / immunology
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • HL-60 Cells
  • Humans
  • Ligands
  • Membrane Glycoproteins / immunology
  • Membrane Glycoproteins / metabolism
  • Mice
  • Molecular Weight
  • P-Selectin / immunology*
  • Platelet Activation / drug effects
  • Rats
  • Rats, Inbred Lew
  • Recombinant Proteins / immunology


  • Antibodies, Blocking
  • Antibodies, Monoclonal
  • Ligands
  • Membrane Glycoproteins
  • P-Selectin
  • P-selectin ligand protein
  • Recombinant Proteins