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Development and characterization of a competitive polyclonal antibody enzyme-immunoassay for salmon insulin-like growth factor-II
Citation
Wilkinson, Ryan and Elliott, P and Hohmann, R and Francis, G and Carragher, J, Development and characterization of a competitive polyclonal antibody enzyme-immunoassay for salmon insulin-like growth factor-II, Comparative Biochemistry and Physiology. Part B: Biochemistry and Molecular Biology, 139, (2) pp. 193-201. ISSN 1096-4959 (2004) [Refereed Article]
DOI: doi:10.1016/j.cbpc.2004.06.016
Abstract
This paper describes the development and validation of a competitive, polyclonal antibody enzyme-immunoassay (EIA) for the measurement of salmon and trout insulin-like growth factor-II (IGF-II). A polyclonal antiserum was raised against a synthetic peptide epitope, corresponding to amino acid residues 1-9 of the N-terminus of mature Atlantic salmon (Salmo salar) IGF-II. The antiserum was purified by hydrophobic charge induction chromatography (HCIC). The partially purified immunoglobulins were used in an enzyme-immunoassay system (EIA) resulting in a highly specific assay for salmon IGF-II with cross-reactivity of less than 0.01% for recombinant salmon IGF-I and recombinant salmon growth hormone (GH), and 5.57% for salmon insulin (sIns). The recombinant salmon IGF-II (rsIGF-II) standard curve limit of detection was 1.37 ng/ml with an EC50 of 44.97±0.82 ng/ml. Intra- and interassay coefficients of variation were determined at 7.47% (n=15) and 7.42% (n=15), respectively. Added rsIGF-II was adequately recovered from acid-treated Atlantic salmon and rainbow trout (Oncorhynchus mykiss) plasma samples. Parallel dose-response inhibition curves were demonstrated for the plasma of both fish species tested. Circulating IGF-II levels of 22.26±2.66 and 18.24±1.43 ng/ml were determined for acid-treated plasma of normal adult Atlantic salmon and rainbow trout, respectively. This EIA should prove to be useful in the study of factors which influence circulating plasma levels of IGF-II in these fish species.
Item Details
| Item Type: | Refereed Article |
|---|---|
| Research Division: | Biological Sciences |
| Research Group: | Biochemistry and cell biology |
| Research Field: | Biochemistry and cell biology not elsewhere classified |
| Objective Division: | Animal Production and Animal Primary Products |
| Objective Group: | Fisheries - aquaculture |
| Objective Field: | Aquaculture fin fish (excl. tuna) |
| UTAS Author: | Wilkinson, Ryan (Dr Ryan Wilkinson) |
| ID Code: | 63067 |
| Year Published: | 2004 |
| Web of Science® Times Cited: | 7 |
| Deposited By: | NC Marine Conservation and Resource Sustainability |
| Deposited On: | 2010-04-13 |
| Last Modified: | 2010-05-04 |
| Downloads: | 0 |
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