Albrethsen Jakob, Bøgebo Rikke, Olsen Jesper, Raskov Hans, Gammeltoft Steen
Department of Clinical Biochemistry, Glostrup Hospital, Glostrup, Denmark.
Clin Chem Lab Med. 2006;44(10):1243-52. doi: 10.1515/CCLM.2006.228.
Surface-enhanced laser desorption-ionization time-of-flight (SELDI-TOF) mass spectrometry of human serum is a potential diagnostic tool in human diseases. In the present study, the preanalytical and analytical variation of SELDI-TOF mass spectrometry of serum was assessed in healthy individuals.
Serum and plasma were obtained from healthy human individuals. Protein peaks in human serum and plasma were determined by SELDI-TOF mass spectrometry.
The protein peaks in serum in healthy individuals showed no variation with gender, age, fasting, or diurnal rhythm. The intensity of protein peaks changed significantly during clotting of serum from 30 to 60 min, followed by smaller changes from 60 to 120 min. The intensity of protein peaks changed after 60-min storage of serum at room temperature, whereas no changes were observed for storage on ice. The number of reproducible protein peaks in serum was determined on WCX2, SAX2, and IMAC30 arrays as 29, 34, and 36, respectively. The average coefficient of variation (CV) of the mass value of protein peaks was 0.03%. The intra-assay CV of peak intensity on IMAC30 arrays was 16% (10%-36%, n=8) for 36 peaks, inter-assay CV was 18% (6%-34%, n=4) for 16 peaks, and inter-individual CV was 38% (16%-56%, n=16) for 20 peaks.
The pre-analytical and analytical conditions of SELDI-TOF mass spectrometry of serum have a significant impact on the protein peaks, with the number of peaks low and the assay variation high. Consequently, SELDI-TOF mass spectrometry of serum needs further evaluation before application in clinical diagnostics.
人血清表面增强激光解吸电离飞行时间（SELDI-TOF）质谱分析是人类疾病潜在的诊断工具。在本研究中，对健康个体血清SELDI-TOF质谱分析的分析前和分析变异进行了评估。
从健康个体获取血清和血浆。通过SELDI-TOF质谱分析测定人血清和血浆中的蛋白质峰。
健康个体血清中的蛋白质峰在性别、年龄、禁食或昼夜节律方面无变化。血清凝固30至60分钟期间蛋白质峰强度变化显著，随后60至120分钟变化较小。血清在室温下储存60分钟后蛋白质峰强度发生变化，而在冰上储存未观察到变化。在WCX2、SAX2和IMAC30芯片上测定血清中可重复蛋白质峰的数量分别为29个、34个和36个。蛋白质峰质量值的平均变异系数（CV）为0.03%。IMAC30芯片上36个峰的峰强度批内CV为16%（10%-36%，n=8），16个峰的批间CV为18%（6%-34%，n=4），20个峰的个体间CV为38%（16%-56%，n=16）。
血清SELDI-TOF质谱分析的分析前和分析条件对蛋白质峰有显著影响，峰数量少且分析变异高。因此，血清SELDI-TOF质谱分析在应用于临床诊断前需要进一步评估。
