一种使用带有固定化糖脲试剂的示踪剂对人血浆中的心房利钠肽进行的灵敏放射免疫测定法。

A sensitive radioimmunoassay of atrial natriuretic peptide in human plasma, using a tracer with an immobilized glycouril agent.

作者信息

Rosmalen F M, Tan A C, Tan H S, Benraad T J

机构信息

Department of Experimental and Chemical Endocrinology, St. Radboud Hospital, University of Nijmegen, The Netherlands.

出版信息

Clin Chim Acta. 1987 Jun 15;165(2-3):331-40. doi: 10.1016/0009-8981(87)90178-1.

DOI:10.1016/0009-8981(87)90178-1

PMID:2958178

Abstract

A highly specific and sensitive radioimmunoassay (RIA) for alpha-human atrial natriuretic peptide (hANP[1-28]) in plasma was developed. The assay used a [125I]monoiodotyrosyl-hANP[1-28] tracer, prepared with an immobilized glycouril agent (Protag) and purified by high pressure liquid chromatography (HPLC), and a highly specific antiserum raised against hANP[1-28], coupled to keyhole limpet haemocyanin, in sheep. Plasma was extracted using C-18 Seppak cartridges. A good parallelism was found after dilution prior to extraction of plasma of patients with congestive heart failure (CHF) or of plasma of healthy subjects. Recovery of hANP[1-28] added to plasma was 96%. The limit of detection was 0.8 pg/tube, intra- and inter-assay variation were 9 and 12%, respectively. Mean plasma ANP values in 25 normal persons with a normal salt intake was 26.0 +/- 15.5 (+/- SD) pg/ml. Plasma levels of 18 subjects (7 normals, 11 CHF) were measured using four different antisera after the extraction step. High correlations were found between the values obtained with these four antisera.

摘要

已开发出一种用于检测血浆中α-人心房利钠肽（hANP[1-28]）的高特异性和高灵敏度放射免疫分析（RIA）方法。该分析方法使用了一种[125I]单碘酪氨酸-hANP[1-28]示踪剂，它是用固定化糖脲试剂（Protag）制备并通过高压液相色谱（HPLC）纯化的，还使用了一种在绵羊体内产生的针对hANP[1-28]的高特异性抗血清，该抗血清与钥孔戚血蓝蛋白偶联。血浆使用C-18 SepPak柱进行提取。对充血性心力衰竭（CHF）患者的血浆或健康受试者的血浆在提取前进行稀释后，发现具有良好的平行性。添加到血浆中的hANP[1-28]回收率为96%。检测限为0.8 pg/管，批内和批间变异分别为9%和12%。25名正常盐摄入量的正常人的平均血浆ANP值为26.0±15.5（±标准差）pg/ml。在提取步骤后，使用四种不同的抗血清对18名受试者（7名正常人，11名CHF患者）的血浆水平进行了测量。发现用这四种抗血清获得的值之间具有高度相关性。