摘要
目的应用血清样品直接标记和抗体芯片方法进行抗原和疾病标志物的筛选。方法分别取正常、糖尿病和甲亢患者的血清,采用FMB的ASB600抗体芯片的样品及相关的标记试剂盒,对3μl血清样本进行直接生物素标记。抗体芯片孵育后,用链霉亲合素标记的Cy3荧光显色芯片,经扫描仪检测杂交信号,分析比较芯片内、芯片之间的信号结果重复性,初步筛选疾病样本中与正常样本有差别的蛋白标志物。结果芯片内的两点重复性相关系数R2达到0.95～0.99,芯片重复实验间的相关系数R2达到0.98。对不同样本间的芯片信号进行比较,可以发现与疾病相关的新蛋白分子。结论本研究中使用的ASB600抗体芯片和直接标记荧光检测的实验方法可用于疾病相关的血清分子标志物的识别与寻找。
Objective Using sample direct labeling strategy to establish an antibody array based on serum disease marker screening platform.Methods Serum samples of 2 health people,2 diabetes and 2 hyperthyroidism patients were collected, and 3μl of each sample was directly labeled with biotin using kits from FMB and coupled with ASB600 antibody arrays. After developed with Streptavidin-Cy3 fluorescence,the image were scanned. the data were submitted to analyze the reproducibility withinthe array and between arrays. The discriminations of the contented serum proteins as potential disease markers were screened.Results The intra slide correlation coefficients reached 0.95-0.99 for the duplicated antibody spots, and 0.98 for duplicated arrays. Compared with the different arrays, the diseases related new markers were detected.Conclusion Using the antibody array ASB600with the direct sample labeling method is applicable for serumdisease marker screening. The systemused in this study is reliable and can be used for screening and explorating new markers in patients with serum disease.
出处
《同济大学学报（医学版）》
CAS
2010年第5期25-29,35,共6页
Journal of Tongji University(Medical Science)
基金
国家高技术研究发展计划资助项目(2006AA020704)
上海市科技人才启明星计划资助项目(08QB14027)
关键词
抗体芯片
标记
疾病标志物
血清
antibody array
direct labeling
disease marker
serum
