荧光法测定碱性磷酸酶及其标记物

Fluorometric Measurement of Alkaline Phosphatase and Its Application to Immunoassay for Human Immunoglobulin G

本文研究了以４－甲基伞形酮磷酸酯（４－ＭＵＰ）为底物测定碱性磷酸酶（ＡＬＰ）活性的高灵敏荧光法及其应用于人血清ＩｇＧ的含量测定．通过纯化４－ＭＵＰ，优化分析条件，使方法的灵敏度有较大提高，实用性也得到改善，测定ＡＬＰ的线性范围为１．７×１０－５～１．７×１０－８ＩＵ／ｍＬ，检出限为４．０×１０－９ＩＵ／ｍＬ，本法已成功地应用于血清样品ＡＬＰ的测定．作为一个例子，通过测定免疫标记物ＨｕＩｇＧ－ＡＬＰ的活性，从而测定了ＩｇＧ的含量，ＨｕＩｇＧ的检出限为２．

A highly sensitive fluorometric assay of alkaline phosphate(ALP) and its application to immunoassay are described. 4 Methylumbelliferyl phosphate(4 MUP) was investigated as fluorogetic substrate for ALP. By purifying the 4 MUP and optimizing the test conditions, the sensitivity was greatly improved. The linear range of ALP is 1.7×10 -5 -6.8×10 -8 IU/mL. The method has been applied successfully to the determination of ALP in human serum. As an example, the determination of human IgG was performed. The detection limit for human IgG is 2.48×10 -8 mg/mL.