Tsuchida Sachio, Satoh Mamoru, Umemura Hiroshi, Sogawa Kazuyuki, Takiwaki Masaki, Ishige Takayuki, Miyabayashi Yui, Iwasawa Yuuya, Kobayashi Sohei, Beppu Minako, Nishimura Motoi, Kodera Yoshio, Matsushita Kazuyuki, Nomura Fumio
Division of Clinical Mass Spectrometry, Chiba University Hospital, Chiba, Japan.
Departments of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
Proteomics Clin Appl. 2018 May;12(3):e1700047. doi: 10.1002/prca.201700047. Epub 2018 Mar 2.
Human serum and plasma are often used as clinical specimens in proteomics analyses, and peptidome profiling of human serum is a promising tool for identifying novel disease-associated biomarkers. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is widely used for peptidomic biomarker discovery. Careful sample collection and handling are required as either can have a profound impact on serum peptidome patterns, yet the effects of preanalytical variables on serum peptidome profiles have not been completely elucidated. The present study investigated the effects of preanalytical variables, including storage temperature, duration (up to 12 months), and thawing methods, on MALDI-TOF MS-based serum peptidome patterns.
Aliquots of serum samples were pretreated with weak cation exchanger magnetic beads using an automated ClinProtRobot system and then analyzed by MALDI-TOF MS.
A number of significant differences in peak intensities were observed depending on sample processing variables.
These peaks can be used as sample quality markers to assess the effects of long-term storage on serum peptidome profiles using MALDI-TOF MS.
人血清和血浆常用于蛋白质组学分析的临床样本,人血清的肽组分析是鉴定新型疾病相关生物标志物的一种有前景的工具。基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)广泛用于肽组生物标志物的发现。需要仔细进行样本采集和处理,因为二者都会对血清肽组模式产生深远影响,然而分析前变量对血清肽组图谱的影响尚未完全阐明。本研究调查了分析前变量,包括储存温度、持续时间(长达12个月)和解冻方法,对基于MALDI-TOF MS的血清肽组模式的影响。
使用自动化ClinProtRobot系统用弱阳离子交换磁珠对血清样本等分试样进行预处理,然后通过MALDI-TOF MS进行分析。
根据样本处理变量观察到许多峰强度的显著差异。
这些峰可用作样本质量标志物,以评估使用MALDI-TOF MS长期储存对血清肽组图谱的影响。