Development of surface plasmon resonance mass spectrometry array platform
Development of surface plasmon resonance mass spectrometry array platform
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DOI：
10.1021/ac070608r
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发表时间：
2007-08-01
影响因子：
7.4
通讯作者：
Nedelkov, Dobrin
中科院分区：
文献类型：
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作者：
Nedelkov, Dobrin
蛋白质微阵列是高通量、高含量蛋白质相互作用分析的首选格式。在大多数阵列形式中，报告分子用于蛋白质相互作用的多步检测。在现有的几种无标记检测方法中，表面等离子体共振（SPR）和质谱（MS）是最有希望用于蛋白质微阵列的方法，尽管这两种方法仅偶尔用于高含量蛋白质阵列。这里首次展示了SPR和MS检测在单个高含量蛋白质微阵列上的组合。五种人血浆蛋白的抗体在化学活化的镀金玻璃芯片上以10 × 10的斑点排列。通过芯片整个表面的SPR成像监测蛋白质与其相应抗体的结合。在蛋白质亲和修复后，将芯片用MALDI基质覆盖并进行MS分析，从阵列上的不同点产生蛋白质特异性质谱。SPR-MS双重检测非常适合高含量蛋白质微阵列和全面的蛋白质分析-从蛋白质浓度的定量评估到检测由遗传变异和表达后处理引起的结构蛋白变体。
Protein microarrays are the format of choice for high-throughput, high-content protein interaction analysis. In most of the array formats, reporter molecules are used in multistep detection of the protein interactions. Among the few existing label-free detection approaches, surface plasmon resonance (SPR) and mass spectrometry (MS) stand out as most promising for utilization in protein microarrays, albeit both have been used only sporadically for high-content protein arrays. Shown here for the first time is the combination of SPR and MS detection on a single high-content protein microarray. Antibodies to five human plasma proteins were arrayed in a 10 x 10 spot arrangement on a chemically activated gold-coated glass chip. Binding of proteins to their corresponding antibodies was monitored via SPR imaging across the entire surface of the chip. Following protein affinity retrieval, the chip was overlaid with MALDI matrix and MS analyzed, producing protein-specific mass spectra from distinct spots on the array. The SPR-MS dual detection is well suited for high-content protein microarrays and comprehensive protein analysis-from quantitative assessment of the protein concentration to detection of structural protein variants arising from genetic variations and postexpression processing.