Objective; To explore a standardized profiling of chronic heart failure with qi-yin deficiency blood proteome in sample collection, preparation, storage procedure and detection by magnetic bead separation and matrix assisted laser desorption / ionization time-of-flight ( MALDI-TOF/TOF) mass spectrometry (MS). Methods; Serum and Plasma samples from 18 healthy volunteers and 18 patients with qi-yin deficiency of chronic heart failure were investigated. Magnetic bead separation (IMAC-Cu, WCX) was used for proteome profiling of human blood by MALOI-TOF-MS and the effects on blood sample collection, proteins extraction, the reproducibility of the proteome analysis, the number of freeze -thaw cycles, samples and ratio of sample and matrix were directly analyzed by evaluating the results of mass spectrometry. Results: In the mass range from 1 000 to 10000 Da,65and39signal peaks of proteins were detected in the serum and plasma of chronic heart failure with qi-yin deficiency patients by IMAC-Cu. 75and47signal peaks of proteins were detected by WCX. Coefficient of variance ( CV) of Eight differential detection of same sample was in the range of 6. 65% ~ 20. 62%. More protein peaks were detected in serum than plasma. The effect of separation by WCX was better than IMAC-Cu. There was significant difference between CHF with qi-yin deficiency and healthy group. There had no influence on blood mass patterns in limited freeze-thaw cycles, different ratios of sample and matrix. Conclusion; The mass spectrometry analysis shows that the stability and reproducibility of the whole system is good to meet the CHF TCM syndrome study requirements. This profiling method may provide reference for TCM syndrome blood proteome study.%目的:建立磁珠法结合MALDI-TOF MS进行慢性心衰气阴虚证血液蛋白质组学研究的实验方法.方法:采用MALDI-TOF-MS技术检测不同磁珠分离的患者血液蛋白质谱,从采集、蛋白提取、反复冻融、重复性、基质样品比例等方面分析其影响.结果:血清质谱峰均多于血浆,弱阳离磁珠出峰数量优于铜整合磁珠,气阴虚证患者与健康组比较蛋白表达有明显差异.结论:该方法作为一种高通量、快速分离、制备和鉴定的蛋白质组学技术,整个实验体系稳定、可靠、重复性好,适用于中医证候蛋白质组学的研究.
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