【摘要】：多肽組學是蛋白質組學技術的延伸和擴展,在醫(yī)學和生物學研究中的應用日益廣泛,但是,多肽組鑒定方法的重現(xiàn)性對實驗結果的影響目前尚不清楚。本研究利用納升液相色譜-高分辨質譜對健康人的尿液多肽組進行了7次平行分析,考察圖譜數(shù)目、圖譜利用率、鑒定的肽段數(shù)目、蛋白質數(shù)目、樣品總離子強度和肽段保留時間等指標的變化,以揭示重復實驗之間分析結果的可變性和穩(wěn)定性。7次測定的肽段數(shù)目平均值為208,標準偏差為38;7次結果合并后,得到了歸屬于114個蛋白質的426個肽段,肽段和蛋白質數(shù)目均顯著增加;而35個蛋白質的109個肽段在所有7次實驗中均被檢出,表明多肽組的單次分析結果既具有一定的隨機性,又具有相對的穩(wěn)定性。增加平行實驗次數(shù)會擴大多肽組數(shù)據(jù)集,但測定3次以上后增加幅度減小。相比于肽段,多肽組的結果在蛋白質水平上更為穩(wěn)定,提示利用蛋白質為多肽組的生物標志物更為穩(wěn)健。
[Abstract]:Polypeptide group is an extension and extension of proteomics technology, and has been widely used in medical and biological research. However, the influence of the reproducibility of polypeptide group identification methods on the experimental results is still unclear. In this study, seven parallel analyses of urine polypeptides from healthy people were carried out by using high resolution mass spectrometry (HRMS). The number of maps, the utilization rate, the number of peptide segments identified, the number of proteins, the number of peptides, and the number of proteins were investigated. The changes of total ionic strength and retention time of peptide segments were used to reveal the variability and stability of the analytical results between repeated experiments. The average number of peptide segments determined for 7 times was 208, and the standard deviation was 387 times after the results were combined. 426 peptides belonging to 114 proteins were obtained, and the number of peptides and proteins increased significantly, while 109 peptides of 35 proteins were detected in all 7 experiments. It also has relative stability. Increasing the number of parallel experiments will enlarge the polypeptide data set, but the increase will decrease after more than 3 times of measurement. Compared with peptide, the results of peptide group were more stable at protein level, suggesting that using protein as biomarker of peptide group was more stable.
【作者單位】： 深圳大學生命科學學院深圳市海洋生物資源與生態(tài)環(huán)境重點實驗室;深圳大學生命科學學院深圳市微生物基因工程重點實驗室;吉林大學第二附屬醫(yī)院中心實驗室;
【基金】：深圳市科技項目(JSGG20140703163838793)資助
【分類號】：O657.63;R446.12

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