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 language | English (US) |
|---|---|
| Pages (from-to) | 151-162 |
| Number of pages | 12 |
| Journal | Veterinary Immunology and Immunopathology |
| Volume | 59 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Oct 6 1997 |
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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 journal › Article
}
TY - JOUR
T1 - Harbor seal (Phoca vitulina) C-reactive protein (C-RP)
T2 - Purification, characterization of specific monoclonal antibodies and development of an immuno-assay to measure serum C-RP concentrations
AU - Funke, Christina
AU - King, Donald P.
AU - Brotheridge, Rory M.
AU - Adelung, Dieter
AU - Stott, Jeffrey L
PY - 1997/10/6
Y1 - 1997/10/6
N2 - 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.
AB - 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.
KW - Acute phase
KW - C-reactive protein
KW - Harbor seal
KW - Immunoassay
KW - Inflammation
KW - Monoclonal antibody
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U2 - 10.1016/S0165-2427(97)00059-7
DO - 10.1016/S0165-2427(97)00059-7
M3 - Article
VL - 59
SP - 151
EP - 162
JO - Veterinary Immunology and Immunopathology
JF - Veterinary Immunology and Immunopathology
SN - 0165-2427
IS - 1-2
ER -