纳升液相色谱-质谱分析方法的重现性对尿液多肽组分析结果的影响
发布时间：2018-10-25 13:52
【摘要】：多肽组学是蛋白质组学技术的延伸和扩展,在医学和生物学研究中的应用日益广泛,但是,多肽组鉴定方法的重现性对实验结果的影响目前尚不清楚。本研究利用纳升液相色谱-高分辨质谱对健康人的尿液多肽组进行了7次平行分析,考察图谱数目、图谱利用率、鉴定的肽段数目、蛋白质数目、样品总离子强度和肽段保留时间等指标的变化,以揭示重复实验之间分析结果的可变性和稳定性。7次测定的肽段数目平均值为208,标准偏差为38;7次结果合并后,得到了归属于114个蛋白质的426个肽段,肽段和蛋白质数目均显著增加;而35个蛋白质的109个肽段在所有7次实验中均被检出,表明多肽组的单次分析结果既具有一定的随机性,又具有相对的稳定性。增加平行实验次数会扩大多肽组数据集,但测定3次以上后增加幅度减小。相比于肽段,多肽组的结果在蛋白质水平上更为稳定,提示利用蛋白质为多肽组的生物标志物更为稳健。
[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.
【作者单位】： 深圳大学生命科学学院深圳市海洋生物资源与生态环境重点实验室;深圳大学生命科学学院深圳市微生物基因工程重点实验室;吉林大学第二附属医院中心实验室;
【基金】：深圳市科技项目(JSGG20140703163838793)资助
【分类号】：O657.63;R446.12
本文编号：2293885
[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.
【作者单位】： 深圳大学生命科学学院深圳市海洋生物资源与生态环境重点实验室;深圳大学生命科学学院深圳市微生物基因工程重点实验室;吉林大学第二附属医院中心实验室;
【基金】：深圳市科技项目(JSGG20140703163838793)资助
【分类号】：O657.63;R446.12
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