Optimization of MALDI-TOF MS detection for enhanced sensitivity of affinity-captured proteins spanning a 100 kDa mass range
Optimization of MALDI-TOF MS detection for enhanced sensitivity of affinity-captured proteins spanning a 100 kDa mass range
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DOI：
10.1021/pr0703526
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发表时间：
2007-11-01
影响因子：
4.4
通讯作者：
Malyarenko, Dariya I.
中科院分区：
文献类型：
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作者：
Gatlin-Bunai, Christine L.;Cazares, Lisa H.;Malyarenko, Dariya I.
MALDI-TOF质谱法对复杂生物样品的分析通常仅限于检测低质量蛋白质（或蛋白质片段）峰。通过对仪器参数、数据处理和亲和捕获的样品制备程序的组合优化，我们将MALDI-TOF高灵敏度检测的质量范围扩大了一个数量级。WCX， C3和IMAC磁珠被确定为互补的，最有利于宽质量范围的蛋白质分析。扩展质量范围的关键仪器参数包括TOF检测器的ADC偏置和前置放大器滤波器值的调整。数据处理通过常数和二次降采样的组合改进，在指数基线减法之前，以增加信号峰值的灵敏度。这种蛋白质信号宽质量范围检测的增强将直接有利于需要全线性范围质量检测的质谱表达谱研究。
Analysis of complex biological samples by MALDI-TOF mass spectrometry has been generally limited to the detection of low-mass protein (or protein fragment) peaks. We have extended the mass range of MALDI-TOF high-sensitivity detection by an order of magnitude through the combined optimization of instrument parameters, data processing, and sample preparation procedures for affinity capture. WCX, C3, and IMAC magnetic beads were determined to be complementary and most favorable for broad mass range protein profiling. Key instrument parameters for extending mass range included adjustment of the ADC offset and preamplifier filter values of the TOF detector. Data processing was improved by a combination of constant and quadratic down-sampling, preceded by exponential baseline subtraction, to increase sensitivity of signal peaks. This enhancement in broad mass range detection of protein signals will be of direct benefit in MS expression profiling studies requiring full linear range mass detection.