Suetomi K, Takahama K
Department of Legal Medicine, Miyazaki Medical College, Japan.
Nihon Hoigaku Zasshi. 1995 Feb;49(1):26-32.
A sandwich enzyme immunoassay for cardiac troponin I (caTnI) is a description which can identify injuries to the heart. Bovine cTnI was purified from bovine cardiac muscle, then an antibody against cTnI was prepared. We confirmed by immunoblotting that the anti-cTnI antibody reacted only to proteins extracted from the heart and in addition, only to cTnI among them. A mouse monoclonal anti-cTnI IgG-coated polystyrene ball was incubated with cTnI, and subsequently with affinity-purified rabbit anti-cTnI Fab-peroxidase conjugate. Specifically bound peroxidase activity was assayed by fluorometry. The detection limit was 3 fmol (84 ng) per assay. The cross-reaction of the sandwich enzyme immunoassay in regard to proteins from other organs was investigated. A little cross-reaction was recognizable only when proteins from skeletal muscle were in high concentration, but we supposed that this assay could be applied to identify injuries to the heart. We developed a sandwich enzyme immunoassay for bovine cTnI. However, further studies will be aimed at developing a sandwich enzyme immunoassay for human cTnI.
一种用于检测心肌肌钙蛋白I（caTnI）的夹心酶免疫测定法是一种可识别心脏损伤的方法。从牛心肌中纯化出牛心肌肌钙蛋白I（cTnI），然后制备抗cTnI抗体。我们通过免疫印迹法确认，抗cTnI抗体仅与从心脏提取的蛋白质发生反应，此外，仅与其中的cTnI发生反应。将包被有小鼠单克隆抗cTnI IgG的聚苯乙烯球与cTnI孵育，随后与亲和纯化的兔抗cTnI Fab-过氧化物酶缀合物孵育。通过荧光测定法检测特异性结合的过氧化物酶活性。每次测定的检测限为3飞摩尔（84纳克）。研究了夹心酶免疫测定法对来自其他器官蛋白质的交叉反应。仅当骨骼肌中的蛋白质浓度很高时，才会出现少量可识别的交叉反应，但我们认为该测定法可用于识别心脏损伤。我们开发了一种用于牛cTnI的夹心酶免疫测定法。然而，进一步的研究将致力于开发一种用于人cTnI的夹心酶免疫测定法。
