Harbor seal (Phoca vitulina) C-reactive protein (C-RP): purification, characterization of specific monoclonal antibodies and development of an immuno-assay to measure serum C-RP concentrations

Veterinary Immunology and Immunopathology
C FunkeJ L Stott

Abstract

C-reactive protein (C-RP) was purified from harbor seal (Phoca vitulina) serum by calcium dependant phosphoryl-choline and protein A affinity chromatography. Polyacrylamide gel electrophoresis under reducing conditions revealed a single protein moiety with a molecular weight of approximately 25 kDa. An internal peptide derived from this purified protein 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 homologue. Eight specific monoclonal antibodies (P13, P51, P87, P101, P106, P130, P157 and P219) were raised against this purified protein. All 8 monoclonal antibodies immunoblotted with the 25 kDa C-RP subunit under reducing conditions. A competitive immunoassay was developed identifying elevated C-RP concentrations in harbor seal serum samples with clinical evidence of inflammatory disease. Application of this immunoassay for the measurement C-RP may provide valuable information for the clinical assessment of harbor seal health.

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Citations

Sep 13, 2003·Developmental and Comparative Immunology·Jamie R CartwrightTrevor J Greenhough
Oct 28, 2005·Journal of Zhejiang University. Science. B·E GruysS J Koopmans
Jan 24, 2014·Journal of Zoo and Wildlife Medicine : Official Publication of the American Association of Zoo Veterinarians·Carolyn CrayKristopher L Arheart
Jul 25, 2014·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Ramiro IsazaCarolyn Cray
Dec 18, 2012·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Carolyn CrayGregory D Bossart
Jun 14, 2019·Frontiers in Immunology·Maria Elena Gelain, Federico Bonsembiante
Sep 19, 2017·Journal of Zoo and Wildlife Medicine : Official Publication of the American Association of Zoo Veterinarians·Julie D SheldonNicole I Stacy
May 23, 2019·Frontiers in Immunology·Asmita Pathak, Alok Agrawal
Jan 23, 2021·Journal of Zoo and Wildlife Medicine : Official Publication of the American Association of Zoo Veterinarians·Jennifer E FlowerMichael J Adkesson

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