个体化医疗的现状与未来-微阵列技术.ppt
个体化医疗的现状与未来四.蛋白质微阵列技术,吕林莉 M.D.,Ph.D.东南大学医学院,The Major Techniques in Current Proteomics,Two-dimensional electrophoresisIEF strip separation SDS-PAGE gel separation Mass SpectrometryProtein sequencing Peptide mappingOthersYeast two hybrid assayProtein microarray,传统蛋白质组研究方法,二维凝胶电泳、质谱技术-需要复杂的设备、样品的需求量大，可重复性低，-在低丰度蛋白检测存在缺陷，且对蛋白水解为多肽不易控制，不利于可靠的数据分析-不能用来作为蛋白质生物标记物的验证,Anderson L et al.J Phsiol 563(1):23-60,2005,Protein Microarrays,Theoretical considerations macrospot-microspot,Roger Ekins:ambient analyte theory,Advantages of Microarray,High sensitivityMuch higher throughputMore flexibilityLess sample consumptionQuantitationDirect target detection,Smaller is better,一种蛋白质的所有变异体 一个蛋白质家族的所有成员一条信号通路中的所有蛋白质 一种组织、器官或有机体的所有蛋白质,一个芯片上可以容纳,Different Kinds of Protein Arrays,Antibody Array Antigen Array Ligand Array,Detection by:SELDI MS,fluorescence,SPR,electrochemical,radioactivity,microcantelever,抗体微阵列(antibody microarray),将抗体作为捕获分子固定于载体表面，通过抗原抗体反应，实现对样品中特定蛋白质的平行性、高通量检测。基于既往对疾病的理解，对疾病相关蛋白质进行高通量的靶向分析。,Haab BB.AACR 97th Annual Meeting April 1-5,2006,Applications,Discover protein expression patterns for molecular classification of diseasesScreen molecular targets for drug developmentDistinguish molecular mechanisms of drug actionDefine biomarkers for disease managementMonitor cytokine profiles of biological samples,Key steps in antibody microarray design,the capture molecules:chemically robust,high affinityImmobolization:strong attachment,retention of function the detection systems:sensitive,signal amplified,easy observed,Attachment Methods:,DiffusionAbsorptionnitrocelluloseCovalent CrosslinkingReactive surfacesAffinity AttachmentAffinity tags,Capture molecules Source Technique,mAb Mouse HybridomascFv/Fab diabodies Antibody libraries Phage display,in vitroAffibodies Microorganism Heterologous expressionAptamers Library SELEX/mRNA display,(DNA/RNA/peptide)Receptor ligands Synthetic Combinatorial chemistryAffinity binding agents Recombinant In vitro evolution,Capture molecules,Detection,国内生物芯片市场,上海博星,上海生物芯片,北京博奥,陕西超英生物科技,西安联尔 2002年，国家药品和食品监督管理局批准了5种生物芯片，除江南生物的地中海贫血基因检测芯片外，其他4种都是蛋白芯片：多肿瘤标志物检测系统、丙型肝炎病毒分片段抗体检测试剂盒、幽门螺旋杆菌抗体检测系统、解脲脲原体、人乳头瘤病毒、沙眼衣原体抗体检测系统.,东南大学吴健雄实验室,哈佛大学蛋白质组学中心,
