摘要
【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 optimum conditions for measuring protein molecular weight were studied using matrix assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS）. [Method]Three different matrices, viz., sinapic acid （SA）, α-cyano-4-hydroxycinnamic acid （CHCA） and 2, 5-dihydroxybenzoic acid （DHB） were tested for determination of molecular weight of bovine serum albumin （BSA） and the effects of different sample concentration were observed on quantification of protein samples. [Result] The results demonstrated that 1 μg protein samples and sinapic acid （SA） matrix dissolved in water solvent （containing 0.1% trifluoroacetic acid and 50% acetonitrile to generate the concentration of 10 mg/mL） produced the best quality of MALDI-TOF-MS spectra. The sample required to determine the exact quantity was optimized as 500 ng. [Conclusion]This technique is convenient, rapid, sensitive and reliable for determining protein molecular weight. The optimal combination was determined to obtain optimum mass spectra, necessary for obtaining accurate molecular weight of highly complicated protein samples.
出处
《南方农业学报》
CAS
CSCD
北大核心
2012年第1期14-17,共4页
Journal of Southern Agriculture
基金
Guangxi Natural Science Foundation(2011GXNSFB018028)
关键词
基质辅助激光解吸电离飞行时间质谱
蛋白质
分子量
优化
matrix assisted laser desorption ionization time of flight mass spectrometry
protein
molecular weight
optimization
