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;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
Journal article (JA)
引用
收藏
页码：815 / 822
相关论文
共 50 条
  • [1] MALDI-TOF MS as a label-free approach to rapid inhibitor screening
    Greis, Kenneth D.
    Zhou, Songtao
    Burt, Thomas M.
    Carr, Andrew N.
    Dolan, Elizabeth
    Easwaran, Vijayasurian
    Evdokimov, Artem
    Kawamoto, Richard
    Roesgen, Jeff
    Davis, Gregory F.
    JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, 2006, 17 (06) : 815 - 822
  • [2] Precision enhancement of MALDI-TOF MS using high resolution peak detection and label-free alignment
    Tracy, Maureen B.
    Chen, Haijian
    Weaver, Dennis M.
    Malyarenko, Dariya I.
    Sasinowski, Maciek
    Cazares, Lisa H.
    Drake, Richard R.
    Semmes, O. John
    Tracy, Eugene R.
    Cooke, William E.
    PROTEOMICS, 2008, 8 (08) : 1530 - 1538
  • [3] Rapid screening of marine bacterial symbionts using MALDI-TOF MS
    Livia M. R. Vidal
    Tainá M. Venas
    Aline R. P. Gonçalves
    Hannah K. Mattsson
    Raphael V. P. Silva
    Maria S. Nóbrega
    Gustavo P. R. Azevedo
    Gizele D. Garcia
    Diogo A. Tschoeke
    Verônica V. Vieira
    Fabiano L. Thompson
    Cristiane C. Thompson
    Archives of Microbiology, 2020, 202 : 2329 - 2336
  • [4] Rapid screening of marine bacterial symbionts using MALDI-TOF MS
    Vidal, Livia M. R.
    Venas, Taina M.
    Goncalves, Aline R. P.
    Mattsson, Hannah K.
    Silva, Raphael V. P.
    Nobrega, Maria S.
    Azevedo, Gustavo P. R.
    Garcia, Gizele D.
    Tschoeke, Diogo A.
    Vieira, Veronica V.
    Thompson, Fabiano L.
    Thompson, Cristiane C.
    ARCHIVES OF MICROBIOLOGY, 2020, 202 (08) : 2329 - 2336
  • [5] Probing label-free intracellular quantification of free peptide by MALDI-ToF mass spectrometry
    Rakowska, Paulina D.
    Lamarre, Baptiste
    Ryadnov, Maxim G.
    METHODS, 2014, 68 (02) : 331 - 337
  • [6] Rapid Identification of Mycobacteria by MALDI-TOF MS
    Kohlmann, V.
    Kehrmann, J.
    Mueller, K. -D.
    Buer, J.
    Rettenmeier, A. W.
    Mosel, F.
    INTERNATIONAL JOURNAL OF MEDICAL MICROBIOLOGY, 2012, 302 : 11 - 12
  • [7] Rapid MALDI-TOF MS identification of commercial truffles
    Khalid El Karkouri
    Carine Couderc
    Philippe Decloquement
    Annick Abeille
    Didier Raoult
    Scientific Reports, 9
  • [8] Rapid identification of Streptomyces isolates by MALDI-TOF MS
    Loucif, Lotfi
    Bendjama, Esma
    Gacemi-Kirane, Djamila
    Rolain, Jean-Marc
    MICROBIOLOGICAL RESEARCH, 2014, 169 (12) : 940 - 947
  • [9] Rapid MALDI-TOF MS identification of commercial truffles
    El Karkouri, Khalid
    Couderc, Carine
    Decloquement, Philippe
    Abeille, Annick
    Raoult, Didier
    SCIENTIFIC REPORTS, 2019, 9 (1)
  • [10] A label-free MALDI TOF MS-based method for studying the kinetics and inhibitor screening of the Alzheimer's disease drug target β-secretase
    Machalkova, Marketa
    Schejbal, Jan
    Glatz, Zdenek
    Preisler, Jan
    ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2018, 410 (28) : 7441 - 7448
12345