摘要
目的检测Clinprot系统中2种磁珠对于同种样品的蛋白分选能力,初步探讨磁珠选择的步骤、指标。方法分别同时利用Cu2+螯合(IMAC-Cu)磁珠和弱阳离子(WCX)磁珠检测同样30例血清标本,其中子宫内膜异位症10例,其他良性卵巢肿物10例,健康人10例。比较2种磁珠分选后的蛋白质谱图基本参数、组内一致性和2种磁珠寻找差异的能力。结果①对于该实验样本,WCX磁珠在蛋白质谱图中总峰数量、峰面积均优于Cu2+磁珠(P<0.001)。而在峰高度上,Cu2+有一定优势;②比较3个组的质谱图组内一致性,WCX与IMAC-Cu均有良好的一致性;③初步了解2种磁珠寻找差异的能力,WCX磁珠与IMAC-Cu磁珠均能区分疾病组与正常组,相对WCX磁珠构建的模型更为稳定。结论在Clinprot实验开始前,选择合适的样本量,综合多个衡量指标,结合多个选择步骤评价不同磁珠抓取蛋白的能力,选择一种磁珠扩大样本量,可以为进一步实验中寻找差异蛋白奠定良好基础。就该实验样品,WCX磁珠更利于扩大样本时应用。
Objective To study the methods for selecting a magnetic bead in Clinprot system. Methods Serum samples from 10 patients with endometriosis, 10 healthy volunteers and 10 patients with other overian diseases were analyzed using either immobilized metal-affinity chromatography containing copper ions （IMAC-Cu） or weak cat ion exchange （MB-WCX）. Proteomic spectra were generated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS）.The spectra were analyzed statistically with flexAnalysisTM and Clinprotools software. Proteomic spectra performed with different magnetic beads were compared, including basic parameter of proteomic spectra, the regularity in same group and the capability of finding differential protein. Results ①The performance of peak amounts and peak area by WCX were significantly better than by IMAC-Cu ,while there were higher peak intensities by IMAC-Cu ; ②On the regularity in same group ,there was no significent difference between the WCX and IMAC-Cu ;③WCX was superior on the capability of finding differential protein and establishing steady model. Conclusion This study indicates that the larger sample amounts, various indexes and specific experiment steps will provide advantage in selecting a more appropriate magnetic bead. According to above-mentioned experiment, WCX is an ideal choice for further experiment.
出处
《生物医学工程与临床》
CAS
2008年第3期189-192,共4页
Biomedical Engineering and Clinical Medicine
基金
国家自然科学基金(30772319)
北京大学人民医院研究与发展基金基础课题(RBD2007-10)
北京市科委基金(H030930040230)
