用于测量白三烯和前列腺素的酶联免疫吸附测定法的开发。

Development of enzyme-linked immunosorbent assays for measurement of leukotrienes and prostaglandins.

作者信息

Miller D K, Sadowski S, DeSousa D, Maycock A L, Lombardo D L, Young R N, Hayes E C

出版信息

J Immunol Methods. 1985 Aug 2;81(2):169-85. doi: 10.1016/0022-1759(85)90202-9.

Abstract

Enzyme-linked immunosorbent assays (ELISAs) were developed for the leukotrienes LTC4 and LTB4 and the prostaglandins 6-keto PGF1 alpha and thromboxane (TxB2). In an indirect assay procedure for all 4 eicosanoids a BSA conjugate of the leukotrienes or an ovalbumin conjugate of the prostaglandins was absorbed to polystyrene microtiter plates. Samples containing the respective eicosanoids were incubated in the coated wells with specific rabbit antisera. The wells were then incubated successively with a goat anti-rabbit antibody linked to fluorescein and a rabbit anti-fluorescein antibody linked to alkaline phosphatase. The resultant assays for LTC4, LTB4, 6-keto PGF1 alpha, and TxB2, gave steep, sensitive inhibition curves; IC50s were 0.2, 10, 1, and 0.4 pmol respectively with minimal cross-reactivity to other eicosanoids. The sensitivities and specificities were comparable to those found in the RIA, and the levels determined in this assay correlate well with those determined in non-immunological assays.

摘要

已开发出用于检测白三烯LTC4和LTB4以及前列腺素6-酮-PGF1α和血栓素（TxB2）的酶联免疫吸附测定法（ELISA）。在针对所有4种类二十烷酸的间接测定程序中，白三烯的牛血清白蛋白缀合物或前列腺素的卵清蛋白缀合物被吸附到聚苯乙烯微量滴定板上。将含有相应类二十烷酸的样品与特异性兔抗血清在包被的孔中孵育。然后，孔依次与连接有荧光素的山羊抗兔抗体和连接有碱性磷酸酶的兔抗荧光素抗体孵育。由此得到的针对LTC4、LTB4、6-酮-PGF1α和TxB2的测定给出了陡峭、灵敏的抑制曲线；IC50分别为0.2、10、1和0.4皮摩尔，与其他类二十烷酸的交叉反应最小。其灵敏度和特异性与放射免疫测定法相当，并且该测定法中测定的水平与非免疫测定法中测定的水平相关性良好。