Ultra-High-Throughput Ambient MS: Direct Analysis at 22 Samples per Second by Infrared Matrix-Assisted Laser Desorption Electrospray Ionization Mass Spectrometry
Ultra-High-Throughput Ambient MS: Direct Analysis at 22 Samples per Second by Infrared Matrix-Assisted Laser Desorption Electrospray Ionization Mass Spectrometry
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DOI：
10.1021/acs.analchem.1c04605
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发表时间：
2022-03-29
影响因子：
7.4
通讯作者：
Pan, Jeffrey Y.
中科院分区：
文献类型：
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作者：
Radosevich, Andrew J.;Pu, Fan;Pan, Jeffrey Y.
红外基质辅助激光解吸电喷雾电离（IR-MALDESI）质谱法是一种环境直接采样方法，正在开发用于高通量，无标记，大规模化合物文库的生化筛选。在这里，我们开发了一种超高通量连续motionIR-MALDESI采样方法，能够在384孔微滴板中以每秒22.7个样品的速率获取数据。在最高速度下，井与井之间的分析物携带率小于1%，3亩m1 -羟咪唑仑和12亩M -右旋孤的信号强度相对标准偏差（RSD）分别为11.5%和20.9%。使用异柠檬酸脱氢酶1 （IDH1）酶测定，能够以类似于30s的时间分辨率对384个样品进行平行动力学研究。最后，我们通过在5.54小时内连续测量来自300个微滴板读数的115200个样品，证明了我们方法的重复性和吞吐量，每个新引入的板的rsd小于8.14%。综上所述，这些结果表明IR-MALDESI的样品采集率超过了目前报道的用于高通量化合物筛选的其他直接采样质谱方法。
Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry is an ambient-direct sampling method that is being developed for high-throughput, label-free, biochemical screening of large-scale compound libraries. Here, were port the development of an ultra-high-throughput continuous motionIR-MALDESI sampling approach capable of acquiring data at rates up to22.7 samples per second in a 384-well microtiter plate. At top speed, less than 1% analyte carryover is observed from well-to-well, and signalintensity relative standard deviations (RSD) of 11.5% and 20.9% for 3 mu M1-hydroxymidazolam and 12 mu M dextrorphan, respectively, are achieved.The ability to perform parallel kinetics studies on 384 samples with a similar to 30s time resolution using an isocitrate dehydrogenase 1 (IDH1) enzyme assay is shown. Finally, we demonstrate the repeatability and throughput of our approach by measuring 115200 samples from 300 microtiter plate reads consecutively over 5.54 h with RSDs under 8.14% for each freshly introduced plate. Taken together, these results demonstrate the use of IR-MALDESI at sample acquisition rates that surpass other currently reported direct sampling mass spectrometry approaches used for high-throughput compound screening.