血清蛋白质组高丰度蛋白去除方法比较研究  

Comparative Study on the Methods for Removal of High-Abundant Proteins in Serum Proteome

作　　者：赵佳威 陆澳 赵洋 孟波 叶子弘 ZHAO Jia-wei;LU Ao;ZHAO Yang;MENG Bo;YE Zi-hong(Zhejiang Provincial Key Laboratory of Biometrology and Inspection&Quarantine,College of Life Sciences,China Jiliang University,Hangzhou 310018,China;Technology Innovation Center of Mass Spectrometry for State Market Regulation,Center for Advanced Measurement Science,National Institute of Metrology,Beijing 100029,China)

机构地区：[1]中国计量大学生命科学学院,浙江省生物计量及检验检疫技术重点实验室,浙江杭州310018 [2]中国计量科学研究院前沿计量科学中心,国家市场监管技术创新中心(质谱),北京100029

出　　处：《质谱学报》2024年第4期519-530,I0004,共13页Journal of Chinese Mass Spectrometry Society

基　　金：科技基础资源调查专项(2022FY101202);国家重点研发计划项目(2022YFF0608400);中国计量科学研究院基本业务费重点项目(AKYZD2111)。

摘　　要：基于健康人群(HC)和肝癌人群(HCC)的血清样本,本研究比较了高选择Top14高丰度蛋白去除试剂盒(Top14)、DMB低丰度蛋白富集试剂盒(DMB)2种血清蛋白质组前处理方案的优劣势。对于HC样本,DMB处理组鉴定的蛋白质数量是Top14处理组的2.6倍,是未处理血清原液(Blank)样本的3.9倍;对于HCC样本,DMB处理组鉴定的蛋白质数量是Top14处理组的3.7倍,Blank组的6.2倍。通过分析蛋白组的丰度排序散点图,发现Top14预处理对高丰度蛋白质的去除效果显著;但相较于DMB预处理,低丰度蛋白质的鉴定覆盖率较低。在蛋白质组鉴定频率方面,DMB处理样本中可定量蛋白质数量明显多于Top14处理组和Blank组。Top14处理组和Blank组中鉴定的蛋白质超过50%可在DMB处理组中被鉴定到,且其鉴定蛋白质的KEGG分析结果基本可在DMB方案中被覆盖。在DMB预处理的肝癌血清中鉴定到47个差异蛋白质,而在Top14处理组和Blank组中分别只鉴定到15、17个差异蛋白质。通过对各组差异蛋白质编码基因的KEGG分析,DMB、Top14预处理组和Blank样本的差异蛋白质分别富集到21、5、1条通路,表明DMB预处理更适用于血清样本蛋白质组的分析。基于DMB预处理方法,本研究发现HSP90AB1、SPP1、ACTR3、SNCA、PECAM1和SRC在肝癌血清样本中显著上调,可作为肝癌筛查候选生物标志物。This study evaluated the protein identification capabilities of the Top14 high-abundance protein removal kit(Top14)and the DMB MagicOmics low-abundance enrichment kit(DMB)on serum samples from healthy individuals(HC)and hepatocellular carcinoma(HCC)patients.For HC samples,DMB treatment led to identifying 2.6 times proteins more than Top14 and 3.9 times more than untreated samples.For HCC samples,DMB achieved 3.7 times increase in protein identification over Top14 and 6.2 times increase over untreated samples.Although the Top14 kit was effective in removing high-abundance proteins,it was less proficient in detecting low-abundance proteins when compared to the DMB method.In terms of protein identification frequency,the DMB-treated samples had a significantly higher number of quantifiable proteins than both the Top14 and Blank groups.Over 50%of the proteins identified in the Top14 and Blank groups were also identified in the DMB group,ensuring a comprehensive proteome coverage as evidenced by KEGG analysis.The DMB method significantly outperformed the others in HCC serum samples,identifying 47 differentially expressed(DE)proteins,in contrast to 15 and 17 identified of the Top14 group and untreated samples,respectively,highlighting its superior ability to uncover critical biomarkers for disease analysis.KEGG pathway analysis showed that DE proteins identified by DMB were involved in 21 distinct pathways,significantly more than 5 and 1 pathways identified by Top14 group and untreated samples,demonstrating DMB’s advanced proteomic profiling capability.This study also underscored HSP90AB1,SPP1,ACTR3,SNCA,PECAM1,and SRC proteins increased in HCC serum samples based on DMB method,marking them as promising HCC biomarkers for disease screening.

关 键 词：血清 蛋白质组 液相色谱-串联质谱(LC-MS/MS) 

分 类 号：O657.63[理学—分析化学]