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目的:利用铜螯合磁珠(magnetic beads-based immobilized metal affinity capture Cu)分离低峰度蛋白,以基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF-MS)技术分别检测骨髓增生异常综合征(myelodysplastic syndromes,MDS)患者和正常人血清蛋白质肽链的质量指纹谱,探索用于MDS辅助诊断的客观方法。方法:收集16例MDS患者(MDS组)和10例正常人(正常对照组)的血清蛋白标本,应用铜螯合磁珠进行蛋白分离,通过MALDI-TOF-MS测定标本的蛋白质谱图,对比分析差异蛋白峰,并建立诊断模型。结果:分子质量1.02～10.25ku范围内共检测到信噪比>3的峰146个,经统计分析,MDS组和正常对照组之间差异明显,有显著性差异峰12个(P<0.05),利用快速分类算法建立诊断模型,模型的识别准确率和交叉验证敏感性分别达93.27%和89.94%。各亚型对比分析,各组间区域交叉,组间差异不明显;根据临床分型,将难治性贫血伴幼稚细胞增多型(RAEB型)与其他类型MDS比较,发现RAEB型患者血清中有1个差异低表达蛋白峰,分子质量约1466.57,经鉴定为纤维蛋白原N端片段。结论:MALDI-TOF-MS技术平台可用于寻找MDS相关的血清蛋白质标志物,其蛋白质谱诊断模型可以区分MDS与正常人,有助于MDS的临床辅助诊断。RAEB型患者与其他类型的MDS中存在差异表达蛋白,提示进展型MDS与其他类型MDS有异质性。
OBJECTIVE: To isolate low-kurtosis proteins by magnetic beads-based immobilized metal affinity capture (Cu-based magnetic beads) and to analyze the effect of matrix-assisted laser desorption / ionization time-of-flight mass spectrometry MALDI-TOF-MS) were used to detect the mass fingerprint of serum protein peptide chain in patients with myelodysplastic syndromes (MDS) and normal persons respectively, and to explore objective methods for the diagnosis of MDS. Methods: Serum protein samples from 16 patients with MDS (MDS group) and 10 healthy people (normal control group) were collected for protein separation. Copper chelate magnetic beads were used for protein separation. The protein profiles were determined by MALDI-TOF-MS. Analysis of differential protein peaks, and the establishment of diagnostic models. Results: A total of 146 peaks with signal to noise ratio> 3 were detected in the range of 1.02 ~ 10.25ku. According to statistical analysis, there were significant differences between MDS group and normal control group (P <0.05) , The rapid classification algorithm was used to establish the diagnosis model, and the recognition accuracy and cross-validation sensitivity of the model were 93.27% and 89.94% respectively. According to the clinical classification, compared with other types of MDS, the patients with RAEB type were found to have RAEB-type anemia A differential expression of low protein peak molecular weight of about 1466.57, identified as fibrinogen N-terminal fragment. Conclusion: The MALDI-TOF-MS technology platform can be used to find serum protein markers related to MDS. The protein profiling model can differentiate MDS from normal subjects and is helpful for the clinical diagnosis of MDS. There are differentially expressed proteins in RAEB patients and other types of MDS, suggesting that progressive MDS is heterogeneous with other types of MDS.