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人血浆中活性肾素的直接免疫测定法。

Direct immunometric assay of active renin in human plasma.

作者信息

Ménard J, Guyenne T T, Corvol P, Pau B, Simon D, Roncucci R

机构信息

Inserm U36, Paris, France.

出版信息

J Hypertens Suppl. 1985 Dec;3(3):S275-8.

PMID:2856717
Abstract

None of the methods currently available can detect the small numbers of active renin (AR) molecules present in plasma. Among seven monoclonal antibodies (Ab), two Abs were selected which did not recognize the same epitope and could be used in a sandwich assay. The first monoclonal Ab, 3E8, binds soluble renin (B 50% = 1 x 10(-10) mol/l) and does not inhibit its enzymatic activity. It was coupled to magnetic beads (Magnogel) and was used to trap both active and inactive renin from 250 microliters plasma. The second Ab, 4G1, binds renin (B 50% = 3.5 x 10(-10) mol/l), inhibits its enzymatic activity, and recognizes inactive renin less than AR. It was iodinated and used to detect AR trapped on Magnogel by the first Ab during a 4-h incubation. The assay can detect 16 pg/ml in human plasma and is highly reproducible. The AR level of 15 normotensive subjects, aged 20-45 years, in an upright posture and on a normal sodium intake, was found to be 41 +/- 18 pg/ml (MRC renin standard). The plasmas were trypsin-activated and their total renin levels were measured with the same pair of monoclonal Abs. The mean value of 286 +/- 142 pg/ml is similar to the value obtained by other assay systems which measure total renin with Abs recognizing both active and inactive renin. The direct measurement of AR provides a convenient and standardized method, since the production of the two monoclonal Abs is unlimited.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

目前可用的方法均无法检测出血浆中存在的少量活性肾素(AR)分子。在七种单克隆抗体(Ab)中,选择了两种不识别相同表位且可用于夹心测定的抗体。第一种单克隆抗体3E8与可溶性肾素结合(半数结合浓度B50% = 1×10⁻¹⁰mol/L),且不抑制其酶活性。它与磁珠(Magnogel)偶联,用于从250微升血浆中捕获活性和非活性肾素。第二种抗体4G1与肾素结合(B50% = 3.5×10⁻¹⁰mol/L),抑制其酶活性,且对非活性肾素的识别低于AR。它经碘化处理,用于在4小时孵育期间检测被第一种抗体捕获在Magnogel上的AR。该测定法可检测人血浆中16 pg/ml的AR,且具有高度可重复性。15名年龄在20 - 45岁、直立姿势且正常钠摄入的血压正常受试者的AR水平为41±18 pg/ml(MRC肾素标准)。血浆经胰蛋白酶激活,并用同一对单克隆抗体测量其总肾素水平。286±142 pg/ml的平均值与其他用识别活性和非活性肾素的抗体测量总肾素的测定系统所获得的值相似。AR的直接测量提供了一种方便且标准化的方法,因为这两种单克隆抗体的生产是不受限的。(摘要截短于250词)

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