使用固定在环氧甲基丙烯酸酯上的萤火虫荧光素酶对血小板中的三磷酸腺苷进行连续流动生物发光测定。

Continuous-flow bioluminescent determination of ATP in platelets using firefly luciferase immobilized on epoxy methacrylate.

作者信息

Carrea G, Bovara R, Girotti S, Ferri E, Ghini S, Roda A

机构信息

Istituto di Chimica degli Ormoni, CNR, Milano, Italy.

出版信息

J Biolumin Chemilumin. 1989 Jan-Mar;3(1):7-11. doi: 10.1002/bio.1170030103.

DOI:10.1002/bio.1170030103

PMID:2711891

Abstract

Firefly luciferase was immobilized on epoxy methacrylate beads and used for a continuous-flow assay of ATP extracted from platelets. The immobilized luciferase had a half-life of 3 days at 25 degrees C; there was a 25% recovery of luciferase activity upon immobilization, and ca 50 reactors were made from 1 mg of commercial enzyme. The sensitivity of the assay was 0.3 pmol of ATP, and the response was linear between 1 and 500 pmol of ATP. The ATP content of platelets obtained with the present method correlated well with those obtained using soluble luciferase.

摘要

萤火虫荧光素酶固定在环氧甲基丙烯酸酯珠上，并用于连续流动测定从血小板中提取的ATP。固定化荧光素酶在25℃下的半衰期为3天；固定化后荧光素酶活性的回收率为25%，1mg商业酶可制成约50个反应器。该测定法的灵敏度为0.3pmol ATP，在1至500pmol ATP之间响应呈线性。用本方法获得的血小板ATP含量与使用可溶性荧光素酶获得的结果相关性良好。