An enzyme immunoassay for rat growth hormone: validation and application to the determination of plasma levels and in vitro release.

Journal of immunoassay Pub Date : 1997-11-01 DOI:10.1080/01971529708005826

E Ezan, E Laplante, M T Bluet-Pajot, F Mounier, S Mamas, D Grouselle, J M Grognet, C Kordon

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引用次数: 23

Abstract

A competitive enzyme immunoassay for rat growth hormone (rGH) has been developed using polyclonal anti-rGH antibodies and an acetylcholinesterase (EC 3.1.1.7.) enzymatic tracer coupled covalently with rGH. The assay was performed in 96-well microtiter plates coated with rabbit polyclonal anti-goat immunoglobulin antibodies. Molecular sieve filtration and Western blot analysis revealed a single immunoreactive peak for rat plasma or pituitary extracts. Cross-reactivity with other rat pituitary hormones or human GH was less than 1%. Assay of samples in a concentration range of 0.7 to 69 ng/ml by enzyme immunoassay and radioimmunoassay were well correlated (r = 0.87 and 0.85 respectively for plasma and culture medium samples). Intra- and inter-assay variations in plasma were 4 (n = 24) and 14% (n = 9) respectively. Minimal detectable amounts of rGH were 0.6 ng/ml. A two-site immunometric assay also developed with the same antibodies allowed a detection threshold of 0.25 ng/ml.

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大鼠生长激素的酶免疫测定:血浆水平和体外释放测定的验证和应用。

采用多克隆抗生长激素抗体和与生长激素共价偶联的乙酰胆碱酯酶(EC 3.1.1.7.)示踪剂，建立了大鼠生长激素(rGH)竞争性酶免疫分析法。在包被兔多克隆抗山羊免疫球蛋白抗体的96孔微滴板上进行检测。分子筛过滤和Western blot分析显示大鼠血浆或垂体提取物存在单一免疫反应峰。与其他大鼠垂体激素或人生长激素的交叉反应性小于1%。在0.7 ~ 69 ng/ml浓度范围内的样品，酶免疫测定和放射免疫测定具有良好的相关性(血浆和培养基样品的r分别为0.87和0.85)。血浆组内和组间差异分别为4% (n = 24)和14% (n = 9)。rGH的最低检测量为0.6 ng/ml。用同样的抗体开发了一种双位点免疫测定法，检测阈值为0.25 ng/ml。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Journal of immunoassay

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