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

Christina Funke, Donald P. King, Rory M. Brotheridge, Dieter Adelung, Jeffrey L Stott

Research output: Contribution to journalArticle

23 Citations (Scopus)

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 tiffs 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.

Original languageEnglish (US)
Pages (from-to)151-162
Number of pages12
JournalVeterinary Immunology and Immunopathology
Volume59
Issue number1-2
DOIs
StatePublished - Oct 6 1997

Fingerprint

Phoca
Phoca vitulina
C-reactive protein
C-Reactive Protein
blood proteins
Blood Proteins
monoclonal antibodies
Monoclonal Antibodies
assays
immunoassays
Immunoassay
proteins
Proteins
Protein Sequence Analysis
Protein Subunits
Staphylococcal Protein A
protein subunits
choline
affinity chromatography
Choline

Keywords

  • Acute phase
  • C-reactive protein
  • Harbor seal
  • Immunoassay
  • Inflammation
  • Monoclonal antibody

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Immunology
  • veterinary(all)

Cite this

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. / Funke, Christina; King, Donald P.; Brotheridge, Rory M.; Adelung, Dieter; Stott, Jeffrey L.

In: Veterinary Immunology and Immunopathology, Vol. 59, No. 1-2, 06.10.1997, p. 151-162.

Research output: Contribution to journalArticle

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