Development of discovering technology to quantify the biomarker for minimally invasive diagnostics.

它们在患病时表达的与器官功能障碍相关的蛋白质特征不佳。我们小组开发了一种通用的化学蛋白质组学方法来鉴定生物标记蛋白。这项研究的目的是利用体内生物素化的化学蛋白质组学来鉴定肝损伤后肝窦内皮细胞中存在的蛋白质生物标记物。在肿瘤坏死因子(TNF)/D-(+)-氨基半乳糖(GalN)联合应用诱导大量肝细胞凋亡的小鼠模型上进行了蛋白质组学研究。在体内，通过向小鼠灌流生物素的活性酯衍生物来进行生物素化，这种生物素衍生物能够实现蛋白质的共价修饰。用从肝炎和正常肝脏中纯化的蛋白质分析蛋白质的表达。用链霉亲和素小球从肝脏提取液中纯化生物素化蛋白，用胰酶消化树脂，并进行质谱分析。这些结果表明，肝损伤上表达的正弦形内皮细胞生物标志物反映了肝炎诱导的分子病理生物学和机制。

The proteins they express at the time of disease and which are related to organ dysfunction are poorly characterized. Our group has developed a general chemical proteomics approach for the identification of biomarker proteins. The aim of this study was to identify protein biomarkers present in sinusoidal endothelial cells following hepatic injury using chemical proteomics employing in vivo biotinylation. We performed a proteomic study in a mouse model of massive hepatocyte apoptosis in tumor necrosis factor (TNF)/D-(+)-galactosamine (GalN) animals induced by the combined administration of GalN and TNF. In vivo biotinylation was carried out by perfusing mice with a reactive ester derivative of biotin that enables the covalent modification of proteins. Protein expression was analyzed using purified proteins from hepatitic and normal livers. The biotinylated proteins were purified from liver extracts using streptavidin beads, digested on the resin by trypsin and subjected to mass spectrometry for identification. These results indicate that the sinusoidal endothelial biomarkers expressed on liver injury reflect the molecular pathobiology and the mechanisms of hepatitis induction.