A Systematic Investigation of the Best Buffers for Use in Screening by MALDI-Mass Spectrometry
A Systematic Investigation of the Best Buffers for Use in Screening by MALDI-Mass Spectrometry
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DOI：
10.1177/1087057116681726
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发表时间：
2017-12-01
期刊：
影响因子：
3.1
通讯作者：
Marshall, Peter
中科院分区：
文献类型：
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作者：
Chandler, Jessica;Haslam, Carl;Marshall, Peter
基质辅助激光解吸/电离质谱法（MALDI-MS）为1536和6144测定格式中的生化靶标筛选提供了一种无标记的替代方案，并可能提供更高的灵敏度、重现性和在指定m/z范围内同时检测多个测定组分。离子抑制效应是MS分析报告的主要限制之一。在MALDI-MS筛选中，已经确定掺入测定中的某些生化组分（例如，用于保持酶、盐和其它添加剂的生理条件的缓冲液）诱导对所监测的分析物离子信号的抑制。这种对离子抑制现象知之甚少的现象是筛选界不愿意将其研究转向样品净化减少的MS方法的关键原因。使用乙酰胆碱作为检测底物模拟物，我们已经产生了强大的数据来量化最常用的成分目前筛选测定中的缓冲液（基础缓冲液、附加组分、去污剂、细胞培养基和其它添加剂）与MALDI-MS相容。这里，最合适的缓冲液和组分，沿着它们在限制离子抑制作用方面的鉴定的最佳浓度，提出用于通过MALDI-MS测量的筛选测定。
Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) offers a label-free alternative for the screening of biochemical targets in both 1536- and 6144-assay formats, as well as potentially providing increased sensitivity, reproducibility, and the simultaneous detection of multiple assay components within a specified m/z range. Ion suppression effects are one of the principal limitations reported for MS analysis. Within MALDI-MS screening, it has been identified that certain biochemical components incorporated into the assay (e.g., the buffers used to preserve the physiological conditions of the enzyme, salts, and other additives) induce suppression of the analyte ion signals monitored. This poorly understood phenomenon of ion suppression is a key reason the screening community has been reluctant to shift their investigations toward MS methods with reduced sample cleanup. Using acetylcholine as an assay substrate mimic, we have generated robust data to quantify the degree to which the most highly used components (base buffers, additional components, detergents, cell culture media, and other additives) within current screening assays are compatible with MALDI-MS. Here, the most suitable buffers and components, along with their identified optimal concentrations in terms of limiting ion suppression effects, are proposed for use in screening assays measured by MALDI-MS.