采用激光荧光检测的胰岛素酶联夹心免疫测定法。
Enzyme-linked sandwich immunoassay for insulin using laser fluorimetric detection.
作者信息
Lidofsky S D, Hinsberg W D, Zare R N
出版信息
Proc Natl Acad Sci U S A. 1981 Mar;78(3):1901-5. doi: 10.1073/pnas.78.3.1901.
Abstract
Human serum samples are assayed for insulin by an enzyme-linked sandwich immunoassay. Horseradish peroxidase is used as an enzyme label for antibody, and enzyme activity is measured by means of the fluorogenic substrate, p-hydroxyphenylacetic acid. The product is detected by excitation of fluorescence with the 325-nm line of a continuous-wave helium/cadmium ion laser on line with reverse-phase high-pressure liquid chromatography. The incubation period is 90 min and the limit of detection of insulin is 30 pM, corresponding to 5 microunits/ml. This method correlates highly with radioimmunoassay, with coefficient of correlation r = 0.95.
摘要
采用酶联夹心免疫分析法检测人血清样本中的胰岛素。辣根过氧化物酶用作抗体的酶标记，酶活性通过荧光底物对羟基苯乙酸进行测定。产物通过与反相高压液相色谱联用的连续波氦/镉离子激光器的325nm谱线激发荧光进行检测。孵育期为90分钟，胰岛素的检测限为30pM，相当于5微单位/毫升。该方法与放射免疫分析法高度相关，相关系数r = 0.95。
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