BIOASSAY OF SERUM LUTEINIZING HORMONE (LH) AND CHORIONIC GONADOTROPIN

1975年，我们开发了黄体生成素生物测定术语RICT(大鼠间质 细胞睾酮)用于测量循环中的黄体生成素，增加5倍 比常规放射免疫分析更灵敏。我们现在有了 开发了一种简单、快速、具有相当灵敏度的方法 用于测定血浆、组织提取物或人绒毛膜促性腺激素 孵化介质。这种生物测定使用的是微滴定板和 产品是通过酶联免疫分析程序测量的 使用转移固相。这项手术可以在更短的时间内完成 超过5小时(而不是超过24小时用于RICT)，最低限度 试剂制备。提出的原则和步骤可以是 也可用于测定脑垂体和下丘脑的激素或 任何能刺激细胞释放的蛋白质激素 可以通过本方法测量的产品(即， 类固醇、环核苷酸、促性腺激素)。 我们使用RICT法来评估分泌的生物促黄体生成素活性。 对内源性和低剂量外源性促性腺激素释放激素的反应 普通人。不存在非特异性等离子体效应 黄体生成素生物测定的结果是无法检测到的 促黄体生成素生物活性水平虽低但可测量 10例促性腺激素减退男性的免疫活性。在普通人身上 外源性小剂量(10ug)静脉注射。GnRH管理结果 优先释放生物活性黄体生成素，从而导致 血浆生物免疫活性中位数显著增加 黄体生成素比率。这种模式模仿了内源性黄体生成素的模式 搏动性。黄体生成素生物活性血浆促黄体生成素优先升高 对外源性或内源性GnRH的反应可能反映了 初始分布体积较小，代谢清除较慢 生物促黄体生成素与免疫促黄体生成素比率。我们已经证明了持续的 一种有效的促性腺激素释放激素对生物活性和免疫活性黄体生成素的抑制作用 拮抗剂(N-乙酰-D-pCLPhel，2-D-Trp3-D-Alal0GnRH10) 绝经后妇女，并显示GnRH拮抗剂 与血清蛋白结合强烈，血浆持续时间长 停留时间。这可能解释了观察到的延长持续时间 体内的拮抗作用。

In 1975 we developed a LH bioassay term RICT (rat interstitial cell testosterone) for measurement of circulating LH, 5-fold more sensitive than conventional radioimmunoassay. We have now developed a simplified and rapid method of comparable sensitivity for the measurement of LH/hCG in plasma, tissue extracts or incubation media. This bioassay uses microtiter plates and the product is, measured by an enzyme-linked immunoassay procedure using transfer solid phase. The procedure can be caried out in less than five hours (versus greater than 24 hr. for RICT) with minimal reagent preparation. The principle and steps presented can be used also for the assay of pituitary and hypothalamic hormones or any protein hormone that can stimulate the release of a cell product which can be measured by the present approach (ie. steroid, cyclic nucleotide, gonadotropin). We used RICT assay to assess biological LH activity secreted in response to endogenous and low dose exogenous GnRH, pulses in normal men. The absence of non-specific plasma effects in the LH bioassay was demonstrated by the finding of undetectable levels of LH bioactivity despite low but measurable immunoactivity in 10 hypogonadotropic men. In normal men exogenous low dose (10 mu g) i.v. GnRH administration resulted in preferential release of bioactive LH, with a consequent significant increase in the median plasma bio- to immunoactive LH ratio. This pattern mimicked that of endogenous LH pulsatility. The preferential increase in bioactive plasma LH in response to exogenous or endogenous GnRH might reflect the smaller initial distribution volume and slower metabolic clearance rate of bio versus immuno LH. We have demonstrated a sustained inhibitory actions on bio and immunoactive LH of a potent GnRH antagonist (N-acetyl-D-pCLPhel,2-D-Trp3-D-Alal0GnRH10) in postmenopausal women, and shown that the GnRH antagonist binds avidly to serum proteins and has a prolonged plasma residence time. This may explain the observed extended duration of the antagonist action in vivo.