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本文介绍了高分子量尿激酶放射免疫分析方法的具体程序以及人尿中高分子量尿激酶的测量结果。标记采用Hunter等改进的氯胺T氧化法。碘化前,高分子量尿激酶用二异丙基氟磷酸失活,以避免生物机体中干扰物质带来的不良影响。失活的高分子量尿激酶与1mci~(125)Ⅰ反应2分钟,反应液全部上Sephadex G—50柱层析,经鉴定,标记率为35～40％。高分子量尿激酶放射免疫测量程序为:反应管中加入一定体积的缓冲液,30μ1~(123)Ⅰ标记的高分子量尿激酶(约2×10~4cpm),标准品或待测样品,反应抗体。总体积0.3ml,摇匀,4℃保温24小时,然后加入20μ1SPA菌体试剂,混匀后于37℃保温0.5小时,3500rpm离心10分钟,测量沉淀放射性。用本法测定5例正常成人尿中高分子量尿激酶值为14.6±6.1iu/ml,灵敏度为0.8iu/ml尿。
This article describes the specific procedures for high molecular weight urokinase radioimmunoassay as well as measurements of human urokinase high molecular weight urokinase. Marked by Hunter improved chloramine T oxidation. Prior to iodination, high molecular weight urokinase is inactivated with diisopropylfluorophosphate to avoid adverse effects of interfering substances in the biological body. Inactivated high molecular weight urokinase and 1mci ~ (125) Ⅰ reaction 2 minutes, the reaction solution all on Sephadex G-50 column chromatography, identified, labeling rate of 35 to 40%. High-molecular-weight urokinase radioimmunoassay program: a certain volume of buffer added to the reaction tube, 30μ1 ~ (123) Ⅰ labeled high molecular weight urokinase (about 2 × 10 ~ 4cpm), standard or test sample, the reaction of antibodies . The total volume of 0.3ml, shake, 4 ℃ incubated for 24 hours, then add 20μ1SPA bacteria reagent, after mixing incubated at 37 ℃ 0.5 hours, 3500rpm centrifuged for 10 minutes to measure the precipitation radioactivity. Using this method, 5 cases of normal adult urine high molecular weight urokinase value of 14.6 ± 6.1iu / ml, the sensitivity of 0.8iu / ml urine.