Kim J Q, Cho H I, Kim S I, Lee H K, Hales C N
Department of Clinical Pathology, Seoul National University College of Medicine, Korea.
J Korean Med Sci. 1989 Dec;4(4):171-7. doi: 10.3346/jkms.1989.4.4.171.
The sensitive and specific immunoradiometric assay is described for human proinsulin and its intermediate peptides (65-66 split and 32-33 split proinsulin). We developed a monoclonal antibody-based two-site immunoradiometric assay with use of streptavidin-biotin labeling. The detection limits of the assays lie in the range of 0.5-2.0 pM. In the proinsulin assay proinsulin cross-reacted 66% with 65-66 split proinsulin but not with insulin or 32-33 split proinsulin. In the assay of 65-66 split proinsulin it does not cross-react with insulin, proinsulin or 32-33 split proinsulin. In the 32-33 split proinsulin assay it cross-reacted 84% with proinsulin and 60% with 65-66 split proinsulin. The precision (C.V.) of the assays was less than 15% over the various concentration. The mean concentrations of insulin, proinsulin, 65-66 split proinsulin and 32-33 proinsulin in eight young male subjects in the fasting state were (pM +/- S.E.M.) 20 +/- 3.6, 2.3 +/- 0.3, undetectable (less than 1.0) and 2.1 +/- 0.7 and at the maximum reached during an oral glucose tolerance test, 150 +/- 26, 9.9 +/- 1.4, 3.8 +/- 0.6 and 19.7 +/- 6.0 respectively.
本文描述了一种用于检测人胰岛素原及其中间肽（65-66裂解胰岛素原和32-33裂解胰岛素原）的灵敏且特异的免疫放射分析方法。我们利用链霉亲和素-生物素标记技术开发了一种基于单克隆抗体的双位点免疫放射分析方法。这些分析方法的检测限在0.5-2.0 pM范围内。在胰岛素原分析中，胰岛素原与65-66裂解胰岛素原的交叉反应率为66%，但与胰岛素或32-33裂解胰岛素原无交叉反应。在65-66裂解胰岛素原分析中，它与胰岛素、胰岛素原或32-33裂解胰岛素原均无交叉反应。在32-33裂解胰岛素原分析中，它与胰岛素原的交叉反应率为84%，与65-66裂解胰岛素原的交叉反应率为60%。在不同浓度范围内，这些分析方法的精密度（变异系数）小于15%。8名空腹状态下的年轻男性受试者中，胰岛素、胰岛素原、65-66裂解胰岛素原和32-33裂解胰岛素原的平均浓度（pM±标准误）分别为20±3.6、2.3±0.3、未检测到（低于1.0）和2.1±0.7，在口服葡萄糖耐量试验期间达到最大值时，分别为150±26、9.9±1.4、3.8±0.6和19.7±6.0。
