Liquid Atmospheric Pressure Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Adds Enhanced Functionalities to MALDI MS Profiling for Disease Diagnostics
Liquid Atmospheric Pressure Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Adds Enhanced Functionalities to MALDI MS Profiling for Disease Diagnostics
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DOI：
10.1021/acsomega.9b01476
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发表时间：
2019-07-01
期刊：
影响因子：
4.1
通讯作者：
Cramer, Rainer
中科院分区：
文献类型：
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作者：
Hale, Oliver J.;Morris, Michael;Cramer, Rainer
液体基质辅助激光解吸/电离（液体MALDI）方法已被开发用于高通量大气压（AP）质谱（MS）分析用于疾病诊断的粗生物液体的分子含量。所提出的方法是快速和高度鲁棒性，使其应用于环境中的速度和低成本的高通量分析是必需的。重要的是，由于产生了多电荷分析物离子，它提供了常规固体MALDI MS分析所缺乏的额外功能，包括使用具有有限m/z范围的高性能质量分析仪。与ESI MS/MS类似，同时实现的上级MS/MS性能为MALDI MS分析增加了更大的分析能力和特异性，同时保留了快速激光分析系统和离线大规模样品制备的优势。这种MALDI MS谱分析方法的潜力在检测奶牛乳腺炎上得到了证明，这是乳制品行业的重大经济负担，每头患病奶牛每年损失数百美元，相当于每年总损失数十亿美元，并且导致使用大量抗生素，增加了抗菌素耐药性的扩散。使用简单的一锅/两步分析物提取，仅制备从每日农场挤奶过程中获得的少量等分试样用于液体MALDI MS分析。使用定制的AP-MALDI离子源进行自动分析，从而能够同时检测脂质、肽和蛋白质。诊断，多电荷，蛋白质离子很容易测序和鉴定的MS/MS实验。使用多变量分析根据乳腺炎状态对样品进行分类，通过“剔除20%”交叉验证确定达到98.5%的准确性（100%特异性）。该方法通常适用于大多数商业高分辨率质谱仪上的AP-MALDI MS分析，并有可能扩展到医院，用于快速评估人类和其他生物流体。
A liquid matrix-assisted laser desorption/ionization (liquid MALDI) method has been developed for high-throughput atmospheric pressure (AP) mass spectrometry (MS) analysis of the molecular content of crude bioliquids for disease diagnostics. The presented method is rapid and highly robust, enabling its application in environments where speed and low-cost high-throughput analyses are required. Importantly, because of the creation of multiply charged analyte ions, it provides additional functionalities that conventional solid MALDI MS profiling is lacking, including the use of high-performance mass analyzers with limited m/z range. The concomitant superior MS/MS performance that is achieved similar to ESI MS/MS adds greater analytical power and specificity to MALDI MS profiling while retaining the advantages of a fast laser-based analysis system and off-line large-scale sample preparation. The potential of this MALDI MS profiling method is demonstrated on the detection of dairy cow mastitis, which is a substantial economic burden on the dairy industry with losses of hundreds of dollars per diseased cow per year, equating to a total annual loss of billions of dollars, as well as leading to the use of large quantities of antibiotics, adding to the proliferation of antimicrobial resistance. Only small amounts of aliquots obtained from the daily farm milking process were prepared for liquid MALDI MS profiling using a simple one-pot/two-step analyte extraction. Automated analysis was performed using a custom-built AP-MALDI ion source, enabling the simultaneous detection of lipids, peptides, and proteins. Diagnostic, multiply charged, proteinaceous ions were easily sequenced and identified by MS/MS experiments. Samples were classified according to mastitis status using multivariate analysis, achieving 98.5% accuracy (100% specificity) determined by "leave 20% out" cross-validation. The methodology is generally applicable to AP-MALDI MS profiling on most commercial high-resolution mass spectrometers, with the potential for expansion into hospitals for rapid assessment of human and other biofluids.