|Budget Amount *help
¥6,200,000 (Direct Cost : ¥6,200,000)
Fiscal Year 1994 : ¥1,300,000 (Direct Cost : ¥1,300,000)
Fiscal Year 1993 : ¥1,500,000 (Direct Cost : ¥1,500,000)
Fiscal Year 1992 : ¥3,400,000 (Direct Cost : ¥3,400,000)
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.
第二段階としてEuを用いた非RI測定法の検討に移った。感度の改善をも狙ってサンドイッチ結合原理による測定法を様々なホルモンについて確立すべく自家製抗体の作製を行い、その結果両生類ゴナドトロピン、ラットPRL、ラットLH、ラットInsulin、ブタLH、FSHに対するウェル第一抗体固相化-Eu標識第一抗体測定系(TR-IFMA、time-resolved immunofluorometric assay)を確立した。現在このIMFAをマルチアッセイに応用すべく検討中である。