MALDI-TOF MS as a Label-Free Approach to Rapid Inhibitor Screening

被引：0
|
作者：
Greis, Kenneth D. [1 ]
Zhou, Songtao [1 ]
Burt, Thomas M. [1 ]
Carr, Andrew N. [1 ]
Dolan, Elizabeth [1 ]
Easwaran, Vijayasurian [1 ]
Evdokimov, Artem [1 ]
Kawamoto, Richard [1 ]
Roesgen, Jeff [1 ]
Davis, Gregory F. [1 ]
机构：
[1] Procter and Gamble Pharmaceuticals, Mason, OH, United States
来源：
Journal of the American Society for Mass Spectrometry | 2006年 / 17卷 / 06期
关键词：
Mass Spectrometry (MS) has been widely reported for measuring the conversion of substrates to products for enzyme assays. These measurements are typically performed by time-consuming LC-MS to eliminate buffer salts that interfere with electrospray ionization MS. However; matrix-assisted laser desorption ionization; time-of-flight MS (MALDI-TOF MS) offers a label-free and direct readout of substrate and product; a fast sampling rate; and is tolerant of many buffer salts; reagents; and compounds that are typically found in enzyme reaction mixtures. In this report; a demonstration of how MALDI-TOF MS can be used to directly measure ratios of substrates and products to produce IC50 curves for rapid enzyme assays and compound screening is provided. Typical reproducibility parameters were <7% RSD-a value comparable to ESI MS quantitative assays and well within the acceptable limits for screening assays. The speed of the MALDI readout is currently about 10 s per sample; thus allowing for over 7500 samples/day. From a simplicity standpoint; the enzymatic reaction mixtures are prepared by liquid handling robots; the reactions are stopped by addition of a 10 times volume of acidic matrix solution; and the samples are simultaneously transferred to MALDI target plate for analysis. Importantly; the ratios of substrate to product are of sufficient reproducibility to eliminate the need for internal standards and; thus; minimize the cost and increasing the speed of assay development. © 2006 American Society for Mass Spectrometry;
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页码：815 / 822
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