Gan Z, Marquardt R R, Xiao H
Department of Animal Science, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada.
Anal Biochem. 1999 Mar 1;268(1):151-6. doi: 10.1006/abio.1998.3053.
A new type of solid-phase assay for proteases and protease inhibitors has been developed using biotinylated casein. The assay involves coating of titer plate wells with biotinylated casein, hydrolysis of this substrate with a protease such as trypsin, reaction of the biotin from the unhydrolyzed substrate with an alkaline phosphatase-streptavidin complex, and finally quantification of the amount of casein remaining on the plate using alkaline phosphatase activity as the indicator. The activity of the bound indicator enzyme is oppositely related to the protease activity of the sample. In addition, the assay can be modified for quantitating the corresponding amount of protease inhibitor in the sample. The assay is simple, sensitive, accurate, inexpensive, and amenable to automation.
一种使用生物素化酪蛋白的新型蛋白酶和蛋白酶抑制剂固相分析方法已被开发出来。该分析方法包括用生物素化酪蛋白包被滴定板孔，用诸如胰蛋白酶的蛋白酶水解此底物，未水解底物中的生物素与碱性磷酸酶-链霉亲和素复合物反应，最后以碱性磷酸酶活性为指标定量测定板上剩余酪蛋白的量。结合的指示酶的活性与样品的蛋白酶活性呈负相关。此外，该分析方法可进行修改以定量样品中相应量的蛋白酶抑制剂。该分析方法简单、灵敏、准确、廉价且适合自动化操作。
