View in EDS

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

Saved in:
Bibliographic Details
Title: MALDI-TOF MS as a Label-Free Approach to Rapid Inhibitor Screening
Authors: Greis, Kenneth D. greis.kd@pg.com, Zhou, Songtao1, Burt, Thomas M.1, Carr, Andrew N.1, Dolan, Elizabeth1, Easwaran, Vijayasurian1, Evdokimov, Artem1, Kawamoto, Richard1, Roesgen, Jeff1, Davis, Gregory F.1
Source: Journal of the American Society for Mass Spectrometry. Jun2006, Vol. 17 Issue 6, p815-822. 8p.
Subjects: Enzymes, Salt deposits, Matrices (Mathematics), Electrospray ionization mass spectrometry
Abstract: 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. [Copyright &y& Elsevier]
Copyright of Journal of the American Society for Mass Spectrometry is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
Description
Abstract: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. [Copyright &y& Elsevier]
ISSN:10440305
DOI:10.1016/j.jasms.2006.02.019