High-Throughput Label-Free Biochemical Assays Using Infrared Matrix-Assisted Desorption Electrospray Ionization Mass Spectrometry
| Autor: | Nathaniel L. Elsen, Sujatha M. Gopalakrishnan, David Chang-Yen, Andrew J. Radosevich, James W. Sawicki, Jon D. Williams, Fan Pu, Nari Talaty |
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| Rok vydání: | 2021 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Desorption electrospray ionization Chromatography Chemistry Diacylglycerol kinase zeta Electrospray ionization 010401 analytical chemistry Ion suppression in liquid chromatography–mass spectrometry 010402 general chemistry Mass spectrometry 01 natural sciences High-Throughput Screening Assays 0104 chemical sciences Analytical Chemistry Matrix (chemical analysis) Matrix-assisted laser desorption/ionization Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Biological Assay |
| Zdroj: | Analytical Chemistry. 93:6792-6800 |
| ISSN: | 1520-6882 0003-2700 |
| Popis: | Mass spectrometry (MS) can provide high sensitivity and specificity for biochemical assays without the requirement of labels, eliminating the risk of assay interference. However, its use had been limited to lower-throughput assays due to the need for chromatography to overcome ion suppression from the sample matrix. Direct analysis without chromatography has the potential for high throughput if sensitivity is sufficient despite the presence of a matrix. Here, we report and demonstrate a novel direct analysis high-throughput MS system based on infrared matrix-assisted desorption electrospray ionization (IR-MALDESI) that has a potential acquisition rate of 33 spectra/s. We show the development of biochemical assays in standard buffers for wild-type isocitrate dehydrogenase 1 (IDH1), diacylglycerol kinase zeta (DGKζ), and p300 histone acetyltransferase (P300) to demonstrate the suitability of this system for a broad range of high-throughput lead discovery assays. A proof-of-concept pilot screen of ∼3k compounds is also shown for IDH1 and compared to a previously reported fluorescence-based assay. We were able to obtain reliable data at a speed amenable for high-throughput screening of large-scale compound libraries. |
| Databáze: | OpenAIRE |
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