蛋白组学定量新方法专题讲座.pptx

报告内容 Outline;背景 Background;甲醛标记引入标记试剂的种类多，标记效率高，但是目前二甲基化定量方法存在定量的动态范围小、定量精密度差等不足 Dimethyl labeling is widely used for proteome quantification due to high labeling efficiency and various reagents, but it suffers from low dynamic range and precision. 由于核结合能的不同导致的质量亏损，通过高分辨率的质谱可以在MS2分辨这微小差异，进而可以在宽线性范围内实现蛋白质组的准确定量 The subtle mass differences, arising from the fact that nuclear binding energy, can be resolved by high MS/MS resolution. High accurate proteome quantification in wide dynamic range can be realized by the paired fragment ions ;实验流程 Workflow;分辨率对定量的影响Effect of MS/MS Resolution on Quantification;定量准确度和精密度Quantification Accuracy and Precision;定量动态范围 Dynamic range ;优势 Advantages;多重标记 Multiplex pIDL ;2. 基于准等重a1离子对的蛋白质组定量方法Pseudo-Isobaric a1 ion pairs for proteome quantification;;定量准确度和覆盖率Quantification accuracy and coverage;;报告内容 Outline;集成化平台 integrated platforms;定量蛋白质组分析的挑战Challenges for proteome quantirfication;17;1.基于SILAC标记集成化定量蛋白质组分析平台SILAC based integrated platform;定量平台性能 Performance of integrated platform;与传统离线方法比较Comparison of integrated platform and off-line protocol;人肝癌高低转移细胞定量蛋白质分析 Quantitative analysis of protein extracts from HCC-P/F cell lines;2.基于18O标记的集成化蛋白质组分析平台18O labeling based integrated platform;固定化酶反应器性能评价Performance of IMER;酶解漏切对定量的影响Effects of missed cleavage digestion on protein quantification;集成化平台中酶反应器残留Carryover on IMER of the integrated platform;定量准确度和精密度 Quantification accuracy and precision;与传统离线方法比较Comparison of integrated platform and traditional off-line method;小鼠腹水淋巴道高低转移细胞蛋白组定量分析 Quantitative analysis of protein extracts from HCA F/P ;3. 基于二甲基化标记的集成化定量分析平台 Integrated platform based on dimethyl labeling ;定量准确度和精密度 Quantification accuracy and precision;在30 h的分析时间内，单次分析可以定量到约2400个蛋白；通过三次分析，最终可以可靠定量到1876个蛋白，其中差异蛋白为79个，其中下调的为29个，上调的为50个。;两种集成化蛋白质组相对定量平台比较Comparison of dimethyl labeling and 18O labeling based integrated platforms;致 谢Acknowledgement