Development of Non-Radioisotopic Immunoassay Systems for Measuring Flounder IGF-I(Endocrinology)
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概要
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A time-resolved fluoroimmunoassay system (TR-FIA) for measuring flounder insulin-like growth factor-I (IGF-I) was developed using biotinylated flounder IGF-I, anti-fish IGF-I antiserum and europium-avidin conjugate. The detection limit per well was <5 pg/well corresponding to <0.5 ng/ml in a basic procedure for sample of 10μl/well and to <0.08 ng/ml in a procedure modified for high volume samples (up to 70 μl/well). Specificity of the assay was validated using various IGF-Is and insulins. All IGFs except seabream IGF-I showed very low or no crossreactivity. Binding inhibition curves for flounder and seabream IGF-Is were completely identical to each other. Intra- and interassay variations ranged from coefficients of variations of 3.9% to 7.2%. Recovery tests using barfin flounder plasma varied from 82.7 to 101.6% in the added range from 20 to 160 ng/ml. This assay system was applied for measuring total plasma IGF-I in barfin flounder injected porcine growth hormone (GH). A group injected with GH at the dose of 0.05 IU/gBW showed a significant increase of total plasma IGF-I compared with those of albumin-injected (control) and initial groups. In addition, I was able to substitute time-resolved fluorometric detection in this assay system with enzymatic fluorometric detection (FIA). Binding inhibition curve for lounder IGF-I in this substituted assay system showed equal performance with that of the TR-FIA system. Correlation of IGF-I levels between TR-FIA and FIA was high (r^2=0.957) in plasma samples from barfin flounders in various physiological conditions. Thus, the present study shows precision and efficiency of two non-radioisotopic immunoassay systems for measuring flounder IGF-I.
- 社団法人日本動物学会の論文
- 2005-09-25
著者
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Andoh Tadashi
Hokkaido National Fisheries Research Institute Fisheries Research Agency
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Andoh Tadashi
Hokkaido National Fisheries Research Institute
関連論文
- Two molecular forms of insulin from barfin flounder, Verasper moseri, are derived from a single gene
- Development of Non-Radioisotopic Immunoassay Systems for Measuring Flounder IGF-I(Endocrinology)
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- PRIMARY STRUCTURES OF INSULINS AND THEIR ENZYME IMMUNOASSAY IN THE BARFIN FLOUNDER, VERASPER MOSERI(PLERONECTIDAE)
- INSULIN RECEPTOR BINDING ACTIVITIES AND THEIR THERMODYNAMICS ANALYZED BY TIME-RESOLVED FLUORORECEPTOR ASSAY IN FISH AND MAMMALS
- Purification and structural determination of insulins, glucagons and somatostatin from stone flounder, Kareius bicoloratus
- INSULIN SECRETION BY ADMINISTRATION OF AMINO ACIDS AND GLUCOSE AND ANABOLIC EFFECT OF AMINO ACIDS AND GLUCOSE BY ADMINISTRATION OF INSULIN IN BARFIN FLOUNDER, VERASPER MOSERI
- ESTABLISHMENT OF NON-RADIOISOTOPIC IMMUNOASSAYS FOR MEASURING PLASMA INSULIN-LIKE GROWTH FACTOR-I (IGF-I) IN BARFIN FLOUNDER (VERASPER MOSERI)
- EFFECTS OF NUTRITIONAL CONDITION ON EXPRESSION LEVEL OF INSULIN GENE AND INSULIN CONCENTRATION IN THE BROCKMANN BODY AND PLASMA INSULIN LEVEL IN BARFIN FLOUNDER, VERASPER MOSERI
- PRIMARY STRUCTURES OF INSULINS FROM BONITO AND GREENLING : AMPLIFICATION OF INSULIN GENE IN FISH
- INSULIN SECRETION ABILITY OF BARFIN FLOUNDER, Verasper moseri