Development of a monoclonal antibody against dentin phosphophoryn: a tool to study odontoblastic activity

The role played by phosphophoryn, one of the major noncollagenous proteins of dentin extracellular matrix, in the mineralization process has not been fully characterized. The purpose of our work was to produce monoclonal antibodies (MAbs) against dentin phosphophoryn and to test their reactivity with primary culture of odontoblasts. Dentin phosphophoryn (DPP) was extracted after the mechanical dissociation of teeth and dialyzed against guanidine and EDTA solutions followed by CaC1(2) precipitation. These extracts were characterized by SDS-PAGE and staining with Coomassie blue and Stains-All. After immunization of mice with these extracts, we produced MAb 7G4, which reacted with dentin phosphophoryn as revealed by Western blot. MAb 7G4 reactivity was tested against a primary culture of pig odontoblasts, revealing filaments specifically stained by the anti-DPP antibody. This antibody will be of great interest to study the mineralization process and dental pulp reaction after capping with various calcium phosphate materials.