DEVELOPMENT OF MULTIPLE-IMMUNOASSAY FOR BIOACTIVE SUBSTANCES.

多重分析是从单个样品中同时测量许多生物活性物质，使得从最少的生物材料中获得许多信息成为可能。我们首先在RIA(放射免疫分析)工作，并成功地使用第一个抗体涂层井连续测量激素。从挽救珍贵的一抗的角度出发，我们又研究了用包被二抗的孔板，建立了大鼠垂体前叶激素的系列放射免疫分析方法。然后，为了建立平行检测，我们用第二抗体包被聚苯乙烯珠子，然后用第一抗体处理它们。我们发现，1/2英寸的小球在串联检测中表现优异，但1/4英寸的小球在平行检测中表现更好。最终建立了多重放射免疫分析方法。用NIDDK试剂盒平行测定大鼠垂体激素组合的结果表明，由于NIDDK TSH标准制剂中存在黄体生成素的污染，因此不能同时测定黄体生成素和促甲状腺激素，其他组合也不会造成任何麻烦。我们进一步研究了利用Eu标记的第一抗体建立多重时间分辨免疫荧光法。到目前为止，我们已经成功地建立了两栖类促性腺激素、大鼠促黄体生成素、大鼠催乳素、大鼠、猪、人胰岛素、猪卵泡刺激素和促黄体生成素的单一RT-IFMA检测方法。

Multiple assay, which measures simultaneously many bioactive substances from a single sample, makes it possible to obtain many information from the least amount of biological materials. We first worked in RIA (radioimmunoassay) , and were successful in measuring hormones serially using first antibody-coated wells. From the standpoint to save the precious first antibodies, we then studied using wells coated with the second antibody, and establidhed serial RIA for rat anterior pituitary hormones. Then, in order to establish parallel assays, we coated polystirene beads with the second antibody, then treated them with the first antibody. We found that 1/2 inch beads were excellent in serial assay, but that 1/4 inch beads were better in parallel assays. Multiple-RIA protocol was finally established. Examination for the combination of rat pituitary hormones for parallel assays using NIDDK kits indicated that LH should not be measured simultaneously with TSH because of the contamination of LH in NIDDK TSH standard preparation, and that other combinations did not make any trouble.We further studied to develop the multiple TR-IFMA (time-resolved immunofluorometric assay) using Eu-labelled first antibody. So far, we succeeded in establishing single TR-IFMA for amphibian gonadotropin, rat LH,rat PRL,rat, porcine, human insulin, porcine FSH and LH.We are now engaged in applying these assay methods to multiple assay.