Urine protein profiling with surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry
Urine protein profiling with surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry
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DOI：
10.1111/j.1523-1755.2004.00352.x
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发表时间：
2004-01-01
影响因子：
19.6
通讯作者：
Nickerson, P
中科院分区：
文献类型：
--
作者：
Schaub, S;Wilkins, J;Nickerson, P
背景资料。在过去的几年里，人们对探索人类蛋白质组越来越感兴趣。特别是，人们的努力集中在开发策略，以生成正常细胞、组织和生物液的可重现的蛋白质图谱，从中研究可以比较不同群体(例如，健康个体与具有特定病理状态的个体)之间的蛋白质表达。使用表面增强激光解吸电离飞行时间质谱仪(SELDI-TOF-MS)分析各种外在因素(仪器设置、基质组成、尿液储存后排空、冻融循环)和内在因素(尿液中的血液、尿液稀释度、首次排尿与中段尿液)对尿蛋白谱的影响。影响尿蛋白图谱重复性和峰值检测的外部因素是基质组成和仪器设置，而冷冻-解冻循环的影响很小。中游尿液样本在4摄氏度下储存三天后，其蛋白质谱没有发生变化。影响正常尿蛋白谱的内在因素是尿液中的血液和尿液稀释度。与中段尿样相比，女性首次排尿中的蛋白质比例有显著不同。当蛋白质组成复杂时，SELDI-TOF-MS技术的局限性包括离子抑制和单个蛋白质的定量。SELDI-TOF-MS为高通量尿蛋白图谱提供了一个独特的平台；然而，分析条件的标准化是至关重要的，必须同时考虑外部和内在因素才能准确地解释数据。
Background. In the last few years there has been an increasing interest in exploring the human proteome. In particular, efforts have focused on developing strategies to generate reproducible protein maps of normal cells, tissues, and biologic fluids, from which studies can then compare protein expression between different groups (e. g., healthy individuals vs. those with a specific pathologic state).Methods. Various extrinsic factors (instrument settings, matrix composition, urine storage post void, freeze- thaw cycles) and intrinsic factors (blood in urine, urine dilution, first- void vs. midstream urine) were analyzed with respect to their impact on urine protein profiling using surface- enhanced laser- desorption/ ionization time- of- flight mass spectrometry (SELDI- TOF- MS).Results. Extrinsic factors that critically influenced reproducibility and peak detection of urine protein profiling were matrix composition and instrument settings, while freeze- thaw cycles had minimal impact. Midstream urines samples did not undergo changes in their protein profile when stored for three days at 4degreesC. Intrinsic factors that influenced normal urine protein profiling were blood in the urine and urine dilution. Female first- void urine had a significantly different ratio of proteins present compared to a midstream urine sample. Limitations of the SELDI- TOF- MS technique included ion suppression and quantification of individual proteins when protein composition was complex.Conclusion. SELDI- TOF- MS offers a unique platform for high throughput urine protein profiling; however, standardization of analysis conditions is critical, and both extrinsic and intrinsic factors must be taken into account for accurate data interpretation.