Radioimmunoassay of canine growth hormone.

Radioimmunoassay of canine growth hormone.
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犬生长激素的放射免疫测定。

DOI:
10.1530/acta.0.0980514
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发表时间:
1981
期刊:
Acta endocrinologica
影响因子:
--
通讯作者:
R. Eigenmann
R. Eigenmann
中科院分区:
--
文献类型:
--
作者:
J. Eigenmann;R. Eigenmann

文献摘要

被引文献

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建立了一种灵敏的犬生长激素(GH)放射免疫分析法。在恒河猴中引发抗体。一种抗血清在1:500 000稀释度下显示出工作滴度。采用乳过氧化物酶酶法进行放射性碘标记。Logit-log转换和最小二乘拟合导致标准曲线在0.39 - 50 ng/ml之间的直线拟合。储存期间形成大分子[125 I]GH导致测定灵敏度降低。因此,通过凝胶色谱法重新纯化[125 I]GH。使用该程序,获得了高的和可重复的测定灵敏度。示踪剂制剂在碘化后使用长达3个月。正常和肢端肥大症犬的稀释血浆导致剂量-反应曲线平行于标准曲线。犬催乳素的交叉反应性为2%。批内变异系数(CV)为3.8,批间CV为7.2%。正常犬的平均血浆GH浓度为1.92 +/- 0.14 ng/ml(平均值+/- SEM)。肢端肥大症犬的GH水平明显较高。胰岛素诱导的低血糖、精氨酸和鸟氨酸给药导致GH增量不一致和缓慢。注射小剂量可乐定可获得更好的反应。可乐定给药hypopolysis犬导致GH增量不存在或较差。
A sensitive radioimmunoassay (RIA) for canine growth hormone (GH) was developed. Antibodies were elicited in rhesus monkeys. One antiserum exhibited a working titer at a dilution of 1: 500 000. Radioiodination was performed enzymatically employing lactoperoxidase. Logit-log transformation and least squares fitting resulted in straight line fitting of the standard curve between 0.39 and 50 ng/ml. Formation of large-molecular [125I]GH during storage caused diminished assay sensitivity. Therefore [125I]GH was re-purified by gel chromatography. Using this procedure, high and reproducible assay sensitivity was obtained. Tracer preparations were used for as long as 3 months after iodination. Diluted plasma from normal and acromegalic dogs resulted in a dose-response curve parallel to the standard curve. Canine prolactin exhibited a cross-reactivity of 2%. The within-assay coefficient of variation (CV) was 3.8 and the between-assay CV was 7.2%. Mean plasma GH concentration in normal dogs was 1.92 +/- 0.14 ng/ml (mean +/- SEM). GH levels in acromegalic dogs were appreciably higher. Insulin-induced hypoglycaemia, arginine and ornithine administration resulted in inconsistent and sluggish GH increment. A better response was obtained by injecting a low dose of clonidine. Clonidine administration to hypopituitary dogs resulted in absent or poor GH increment.