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    • 6. 发明申请
    • High throughput screen for inhibitors of polypeptide aggregation
    • 高通量筛选多肽聚集抑制剂
    • US20070077552A1
    • 2007-04-05
    • US11542564
    • 2006-10-03
    • Michael HechtWoo KimChristine Wurth
    • Michael HechtWoo KimChristine Wurth
    • C12Q1/04C12Q1/00
    • G01N33/6896G01N2500/00
    • We have developed a high through-put screen capable of isolating inhibitors of polypeptide aggregation, such as Alzheimer's Disease polypeptide Aβ aggregation, or other disease state aggregating proteins, from amidst large libraries of candidate inhibitors. The screen uses a fusion of a polypeptide domain that self-aggregates, such as an Aβ42 domain characteristic of Alzheimer's disease plaques, to a reporter construct, such as Green Fluorescent Protein (GFP) or similar fluorescent protein. In the absence of inhibition, the rapid misfolding and aggregation of Aβ42 causes the entire fusion protein to misfold, thereby preventing fluorescence. Compounds that inhibit Aβ42 aggregation enable GFP to fold into its native structure, and can be identified by the resulting fluorescent signal.
    • 我们开发了一种能够从候选抑制剂的大型文库中分离出能够分离多肽聚集抑制剂的高通量筛选,如阿尔茨海默氏病多肽Abeta聚集或其他疾病状态聚集蛋白。 屏幕使用自聚集的多肽结构域(例如阿尔茨海默氏病斑块特征的Abeta42结构域)与报道构建体如绿色荧光蛋白(GFP)或类似荧光蛋白的融合。 在没有抑制作用的情况下,Abeta42的快速错误折叠和聚集导致整个融合蛋白错误折叠,从而防止荧光。 抑制Abeta42聚集的化合物使GFP能够折叠成其天然结构,并且可以由所得到的荧光信号鉴定。