Bodola F, Benedict C R
Division of Cardiology, University of Texas Medical Branch, Galveston 77550.
Clin Chem. 1988 May;34(5):970-3.
We have improved a radioimmunoassay for arginine-vasopressin (AVP) and atrial natriuretic peptide (ANP) by using Sep-Pak C18 cartridges to extract AVP and ANP from acidified plasma. The analytes are co-eluted by use of a mobile phase consisting of 1,2-dimethoxyethane and 40 g/L aqueous trifluoroacetic acid (95/5, by vol). After rapid evaporation of the solvents, AVP and ANP are assayed by a nonequilibrium radioimmunoassay method in which commercially available antibodies and radiolabeled antigens are used. The bound fractions are separated from the free by use of polyethylene glycol with human gamma globulin and rabbit anti-human IgG as the second antibody. This results in very low nonspecific binding: 0.44% for the ANP assay, 0.70% for AVP. The minimum detectable amount of ANP is 0.39 pg per tube; for AVP, it is 0.13 pg per tube. Compared with other published methods, this method is substantially more reliable, economical, and easily established in a clinical chemistry laboratory.
我们改进了一种用于精氨酸加压素（AVP）和心钠素（ANP）的放射免疫分析法，通过使用Sep-Pak C18柱从酸化血浆中提取AVP和ANP。分析物通过使用由1,2-二甲氧基乙烷和40 g/L三氟乙酸水溶液（体积比95/5）组成的流动相进行共洗脱。溶剂快速蒸发后，AVP和ANP通过非平衡放射免疫分析法进行检测，该方法使用市售抗体和放射性标记抗原。结合部分通过使用聚乙二醇与人γ球蛋白和兔抗人IgG作为第二抗体从游离部分中分离出来。这导致非常低的非特异性结合：ANP分析为0.44%，AVP为0.70%。ANP的最低检测量为每管0.39 pg；对于AVP，为每管0.13 pg。与其他已发表的方法相比，该方法在临床化学实验室中更可靠、经济且易于建立。
