Biomarkers probed in saliva by surface plasmon resonance imaging coupled to matrix-assisted laser desorption/ionization mass spectrometry in array format
Biomarkers probed in saliva by surface plasmon resonance imaging coupled to matrix-assisted laser desorption/ionization mass spectrometry in array format
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DOI：
10.1007/s00216-014-8373-8
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发表时间：
2015-02
影响因子：
4.3
通讯作者：
Johana Musso;W. Buchmann;F. Gonnet;N. Jarroux;S. Bellon;C. Frydman;Didier-Luc Brunet;R. Daniel
中科院分区：
文献类型：
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作者：
Johana Musso;W. Buchmann;F. Gonnet;N. Jarroux;S. Bellon;C. Frydman;Didier-Luc Brunet;R. Daniel
蛋白质生物标记物的检测是蛋白质组学的主要研究方向。本研究报告了通过表面等离子体共振成像耦合质谱（SPRi-MS）直接从生物流体（人类唾液）中分析蛋白质生物标志物，使用阵列格式的功能化生物芯片进行多重SPRi-MS分析。SPR生物芯片的金表面由自组装的带有ann -羟基琥珀酰亚胺端基的短聚环氧乙烷链单层功能化，用于固定抗体。实验在没有任何样品预纯化或用目标生物标记物进行标记的情况下完成。通过SPRi监测相互作用，从生物芯片表面进行免疫捕获，最后在芯片上基质辅助激光解吸/电离-质谱法对唾液α-淀粉酶和溶菌酶两种蛋白质生物标志物进行结构鉴定。对于溶菌酶，芯片上的MS鉴定是通过基于芯片上蛋白水解程序和肽质量指纹图谱的蛋白质组学分析完成的。
Detection of protein biomarkers is of major interest in proteomics. This work reports the analysis of protein biomarkers directly from a biological fluid, human saliva, by surface plasmon resonance imaging coupled to mass spectrometry (SPRi-MS), using a functionalized biochip in an array format enabling multiplex SPR-MS analysis. The SPR biochip presented a gold surface functionalized by a self-assembled monolayer of short poly(ethylene oxide) chains carrying anN-hydroxysuccinimide end-group for the immobilization of antibodies. The experiments were accomplished without any sample pre-purification or spiking with the targeted biomarkers. SPRi monitoring of the interactions, immune capture from the biochip surface, and finally on-chip matrix-assisted laser desorption/ionization-MS structural identification of two protein biomarkers, salivary α-amylase and lysozyme, were successively achieved directly from saliva at the femtomole level. For lysozyme, the on-chip MS identification was completed by a proteomic analysis based on an on-chip proteolysis procedure and a peptide mass fingerprint.