利用MALDI-TOFMS进行恶性肿瘤血清蛋白组学研究方法的建立

Establishment of standard methods in cancer serum proteomics research with MALDI-TOF MS

目的:建立一套稳定的利用MALDI-TOF MS进行恶性肿瘤血清蛋白质组学分析方法。方法:通过对血清样品采集、保存条件的优化,标准蛋白质的选择,样品的前处理和上样分析等多种条件的优化,确定各个相关参数的选择。结果:经实验条件的比较与优化发现,血液样本采集1 h后经2000 g离心分离血清,分装成若干份,冻存于-80℃冰箱备用,样本前处理方法参照说明书进行;质谱分析条件为应用线性模式,采集范围800-10,000Da,利用多点采集的方法,8点采集,累加至400shots,保存备用。结论:通过优化实验条件,初步建立了利用MALDI-TOF MS进行血清蛋白质组学分析的一整套研究方法,生物质谱分析结果的整体稳定性和重复性可满足统计学和临床医学实验室分析要求。

Objective：To establish standard methods to study cancer serum proteomics with MALDI-TOF MS.Methods：We optimized the selection of blood collection tubes,the serum samples collection,conservation condition,pre-extraction procedure,standard protein and peptides chosen and MALDI-TOF MS reference parameters setting.Results：The best conservation condition was to put numbers on the tubes at first,then clotting at least one hour before centrifugation and put to-80℃ for up to a year,using microflex MS,linear mode,mass range 800-10,000Da,accumulate 8 spots and add up to 400shots to save.Conclusion：By optimizion,established standard methods used in biochemistry laboratory,the mass spectrometry analysis shows that the stability and reproducibility is good to meet the statistical analysis and clinical medicine laboratory requirements.So we can do more samples analysis by using these methods,and the proteomics information is stable and reproductive.