Sanghera Jasbinder, Li Rick, Yan Jun
SignalChem Inc., Richmond, British Columbia, Canada.
Assay Drug Dev Technol. 2009 Dec;7(6):615-22. doi: 10.1089/adt.2009.0237.
Many assay technologies have been developed and utilized to efficiently assay and screen against protein kinase targets. The radiometric assay format for assaying the protein kinase targets has been considered the "Gold Standard" format since it allows the direct readout of kinase functional activity and is a universal assay that is highly sensitive. However, the hazardous nature of the radiometric assay together with the regulatory hurdles has led to the development of alternative assay formats for assessing protein kinase activity measurements. The luminescent ADP-Glo assay has been developed as an alternative to radiometric format for assaying protein kinase targets. This assay allows the measurement of the ADP product formed during the kinase reaction. Therefore, the luminescent ADP-Glo assay is similar to the radiometric format in that it measures the direct product of the protein kinase reaction. Furthermore, since the ADP product is generated by all protein kinase reactions, this is a universal format that can be used for assaying any given protein kinase target. Analysis of data generated with multiple protein kinase targets and the luminescent ADP-Glo technology shows comparable results to the radiometric assay format. Therefore, the luminescent ADP-Glo assay is a robust new technology for evaluating catalytic function of protein kinases as well as other ATPases.
已经开发并利用了许多检测技术来有效地检测和筛选蛋白激酶靶点。用于检测蛋白激酶靶点的放射性检测方法一直被视为“金标准”方法,因为它可以直接读出激酶的功能活性,并且是一种高度灵敏的通用检测方法。然而,放射性检测的危险性以及监管障碍促使人们开发替代检测方法来评估蛋白激酶活性测量。发光ADP-Glo检测方法已被开发出来,作为检测蛋白激酶靶点的放射性检测方法的替代方法。该检测方法可以测量激酶反应过程中形成的ADP产物。因此,发光ADP-Glo检测方法与放射性检测方法类似,因为它测量的是蛋白激酶反应的直接产物。此外,由于ADP产物是由所有蛋白激酶反应产生的,这是一种通用方法,可用于检测任何给定的蛋白激酶靶点。对多个蛋白激酶靶点和发光ADP-Glo技术产生的数据进行分析,结果显示与放射性检测方法相当。因此,发光ADP-Glo检测方法是一种用于评估蛋白激酶以及其他ATP酶催化功能的强大新技术。