免疫学技术-2新技术应用201212.ppt

原理 对蛋白质溶液样品进行各种类型的同核或异核多维核磁共振，建立三维结构数据库。 精度 可与2?分辨率的X射线衍射晶体结构分析相比较. Multiplexed Particle-based Flow Cytometric Assays Variant antigenic peptide promotes cytotoxic T lymphocyteadhesion to target cells without cytotoxicit ( time-lapse video microscopy ) ◆ DNA sequences ◆ Protein structures ◆ Capillary flow data ◆ Chemical structures ◆ Chemical synthesis products ◆ Genetic mutations ◆ Medical data 计算机图形工作站：进行蛋白质分子结构研究的前提 1、运算系统:CPU,主板控制芯片,内存 2、存储设备：硬盘、光盘 3、图形处理器：图形加速卡，显示器 4、操作系统：UNIX、WINDOWS NT、LINUX Nature Immunology 3, 887 - 889 (2002) Frontiers in peptide¨CMHC class II multimer technology In site tetramer 原位四聚体技术 A：SIV A *01/Gag tetramer 染色 B： 抗CD8抗体染色 C： A＋ B Ig gene transcripts showed in different types of cancer cells By Single Cell RT-PCR Detection of Ig gene rearrangement of V-D-J(heavy chain) by RT-PCR V D J C RT-PCR V D J C γ/μ chain Detection of Ig gene rearrangement of V-J(kappa chain) by RT-PCR V J C RT-PCR V J C Kappa chain ◆ 激光捕获显微分离(Laser Capture Microdissection －LCM ????) 整体成像技术 Zhao L，Ding YQ et al. Gut 59:1226-1235, 2010 LIM and SH3 protein 1(LASP-1) 促进肿瘤生长与转移 Pan QJ et al. Internal Immunopharmacology. 10 (2010) 967–977 体内成像技术检测LPS/D-GalN诱发致死性肝损伤的NF-kB activity 活性 Stabilized imaging of immune surveillance in the mouse lung Mark R Looney et al Nature methods 8 : 91-98, 2011 实时细胞内成像技术 芯片技术 ◆ 基因芯片 ◆ 蛋白芯片 ◆ 肽芯片 ◆ 糖芯片 ◆ 抗体芯片 ◆ 液体芯片 …… Lymphochip Clustering Analysis Defined Gene Expression Signatures Gene Expression Physiology of Immune Response Antibody-based Protein Array 1. Incubate soluble sample on plate or  membrane-immobilized antibody array 2. Add a mix of labeled soluble antibody  against the same set of antigens 3. Incubate with developing system and quantitate signal Full set of data 1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6 Nature Biotechnology 20:270, 2002 Nature Biotechnology, 20:275,