個體化醫(yī)療的現(xiàn)狀與未來微陣列技術

1、個體化醫(yī)療的現(xiàn)狀與未來四. 蛋白質微陣列技術,呂林莉 M.D., Ph.D.東南大學醫(yī)學院,The Major Techniques in Current Proteomics,Two-dimensional electrophoresisIEF strip separation SDS-PAGE gel separation Mass SpectrometryProtein sequencing Peptide ma

2、ppingOthersYeast two hybrid assayProtein microarray,傳統(tǒng)蛋白質組研究方法,二維凝膠電泳、質譜技術 ---需要復雜的設備、樣品的需求量大，可重復性低， ---在低豐度蛋白檢測存在缺陷，且對蛋白水解為多肽不易控制，不利于可靠的數(shù)據(jù)分析 ---不能用來作為蛋白質生物標記物的驗證,Anderson L et al. J Phsiol 563(1) :23-60,2005,P

3、rotein Microarrays,,,Theoretical considerations macrospot-microspot,Roger Ekins: ambient analyte theory,Advantages of Microarray,High sensitivityMuch higher throughputMore flex

4、ibilityLess sample consumptionQuantitationDirect target detection,Smaller is better,一種蛋白質的所有變異體 一個蛋白質家族的所有成員一條信號通路中的所有蛋白質 一種組織、器官或有機體的所有蛋白質,一個芯片上可以容納,Different Kinds of Protein Arrays,,,,,,,,,,,,,,,,,,,,,,,,,Antibo

5、dy Array Antigen Array Ligand Array,Detection by: SELDI MS, fluorescence, SPR, electrochemical, radioactivity, microcantelever,抗體微陣列(antibody microarray),將抗體作為捕獲分子固定于載體表面，通過抗原抗體反應，實現(xiàn)對樣品中特定蛋白質的平行性、高通量檢測。

6、基于既往對疾病的理解，對疾病相關蛋白質進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 development

7、Distinguish 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, h

8、igh affinityImmobolization: strong attachment, retention of function the detection systems: sensitive, signal amplified, easy observed,Attachment Methods:,DiffusionAbsorptionnitrocelluloseCovalent CrosslinkingRe

9、active surfacesAffinity AttachmentAffinity tags,Capture molecules Source Technique,mAb Mouse HybridomascFv/Fab diabodies Antibody libraries

10、 Phage display, in vitroAffibodies Microorganism Heterologous expressionAptamers Library SELEX/mRNA display,(DNA/RNA/peptid

11、e)Receptor ligands Synthetic Combinatorial chemistryAffinity binding agents Recombinant In vitro evolution,,Capture molecules,Detection,,,,國內生物芯片市場,上海博星, 上海生物芯片,北