HARBOR SEAL (PHOCA-VITULINA) C-REACTIVE PROTEIN (C-RP) - PURIFICATION, CHARACTERIZATION OF SPECIFIC MONOCLONAL-ANTIBODIES AND DEVELOPMENT OF AN IMMUNOASSAY TO MEASURE SERUM C-RP CONCENTRATIONS

C-reactive protein (C-RP) was purified from harbor seal (Phoca vitulin a) serum by calcium dependant phosphoryl-choline and protein A affinit y chromatography. Polyacrylamide gel electrophoresis under reducing co nditions revealed a single protein moiety with a molecular weight of a pproximately 25 kDa. An internal peptide derived from this purified pr otein was subjected to N-terminal amino acid sequencing. A high amino acid sequence similarity was obtained with other published mammalian C -RP molecules confirming that the purified protein was a C-RP homologu e. Eight specific monoclonal antibodies (P13, P51, P87, P101, P106, P1 30, P157 and P219) were raised against this purified protein. All 8 mo noclonal antibodies immunoblotted with the 25 kDa C-RP subunit under r educing conditions. A competitive immunoassay was developed, identifyi ng elevated C-RP concentrations in harbor seal serum samples with clin ical evidence of inflammatory disease. Application of this immunoassay for the measurement C-RP may provide valuable information for the cli nical assessment of harbor seal health. (C) 1997 Elsevier Science B.V.