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【目的】研究采用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术检测蛋白质分子量的优化条件。【方法】选择3种不同的基质检测牛血清白蛋白(BSA)的分子量,在选定最佳基质化合物的基础上,通过改变样品蛋白质的浓度,以获得高质量的质谱峰图,确定蛋白质样品的最小和最优检测量。【结果】1μg蛋白质样品与10mg/mL含0.1%乙酸(TFA)和50%乙腈(ACN)的芥子酸(SA)基质水溶液混合检测分子量,可产生最高质量的MALDI-TOF-MS峰图,同时蛋白质样品最小检测量应不少于500ng。【结论】MALDI-TOF-MS技术操作简单、分析快速、灵敏度高,是一种检测蛋白质分子量的可靠方法;研究结果确定了获得最优化质谱峰的最佳组合,这是从高度复杂的蛋白质样品测定准确分子量的必备条件。
【Objective】 The objective of this study was to investigate the optimum conditions for the determination of protein molecular weight by matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS). 【Method】 The molecular weight of bovine serum albumin (BSA) was determined by selecting three different substrates. Based on the selected optimal matrix compounds, high quality mass spectra were obtained by changing the protein concentration of the samples, and the protein samples The minimum and optimal detection volume. [Results] The highest quality MALDI-TOF-MS peak pattern was generated when 1 μg protein sample was mixed with 10 mg / mL aqueous solution of erucic acid (SA) matrix containing 0.1% acetic acid (TFA) and 50% acetonitrile (ACN) The minimum amount of protein sample should be at least 500ng. 【Conclusion】 MALDI-TOF-MS is a simple, rapid and sensitive method for detecting protein molecular weight. The results of the study confirm the optimal combination for obtaining the most optimized mass spectral peak, which is obtained from highly complex protein samples Prerequisites for determining the exact molecular weight.