Radioimmunoassay of human high molecular weight kininogen in normal and deficient plasmas.
Radioimmunoassay of human high molecular weight kininogen in normal and deficient plasmas.
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正常和缺陷血浆中人高分子量激肽原的放射免疫测定。
DOI：
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发表时间：
1980
期刊：
影响因子：
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通讯作者：
J. Pisano
中科院分区：
文献类型：
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作者：
D. Proud;J. Pierce;J. Pisano
本文报道了一种测定人高分子量激肽原的放射免疫分析法，可检测150 pg的抗原。用免疫亲和层析法纯化高分子量激肽原抗体，用双抗体沉淀法分离游离抗原和结合抗原。在LMW激肽原中，仅一种检测形式（B3.2）显示出显著的交叉反应（2%）。缓激肽和人血浆激肽释放酶均未显示交叉反应，猴HMW激肽原显示与人抗原相同。批内和批间变异系数分别为2%和1.5%。加入6份血浆的HMW激肽原回收率为97.7% +/-1.8%。17份正常血浆的测定结果为90.8 ± 2.5 μ g/ml HMW激肽原（平均值± S.E.M.）。基于纯化血浆激肽释放酶对激肽的特异性释放，对样品进行生物测定，得到90.2 +/- 2.8 μ g/ml HMW激肽原水平（r = 0.83，p小于0.001）。在两种试验中均未观察到任何显著的性别差异。在正常血浆的凝胶过滤后未观察到任何抗原片段的证据。RIA测量也进行了7个血浆报告缺乏HMW激肽原。威廉姆斯、代顿、旧金山弗朗西斯科和福劳耶克血浆均未显示出显著的交叉反应，而菲茨杰拉德、里德和底特律血浆分别显示出正常抗原水平的1.0%、2.5%和3.5%。这种灵敏、方便的方法将有助于研究激肽释放酶-激肽系统在健康和疾病中的作用。
An RIA for human HMW kininogen, capable of detecting 150 pg of antigen, has been developed. Antibody to HMW kininogen was purified by immunoaffinity chromatography, and double-antibody precipitation was used to separate free and bound antigen. Of the LMW kininogens only one of the forms tested (B3.2) showed significant cross-reaction (2%). Bradykinin and human plasma kallikrein both showed no cross-reaction, and monkey HMW kininogen showed identity to the human antigen. Intraassay and interassay coefficients of variation were 2% and 1.5%, respectively. Recovery of HMW kininogen added to 6 plasmas was 97.7% +/- 1.8%. Assay of 17 normal plasmas gave a level of 90.8 +/- 2.5 micrograms/ml HMW kininogen (mean +/- S.E.M.). A bioassay of the samples, based on specific release of kinin by purified plasma kallikrein, yielded a level of 90.2 +/- 2.8 micrograms/ml HMW kininogen (r = 0.83, p less than 0.001). In neither assay was any significant sex difference observed. No evidence of any antigenic fragments was seen upon gel filtration of normal plasmas. RIA measurements were also performed on seven plasmas reportedly deficient in HMW kininogen. Williams, Dayton, San Francisco, and Flaujeac plasmas all showed no significant cross-reaction, whereas Fitzgerald, Reid and Detroit plasmas showed 1.0%, 2.5%, and 3.5% of normal antigenic levels, respectively. This sensitive, convenient method should facilitate studies on the role of the kallikrein-kinin system in health and disease.