Multiplexed HTS of Serine and Cysteine Proteases (RMI)

描述（由申请人提供）：针对与凝血、补体功能、中性粒细胞血小板串扰和溶酶体过程相关的一系列蛋白质，提出了多重微阵列HTS活动。定义了将化合物文库以重复组打印在微阵列上的方案。每组微阵列（3072种化合物/微阵列和33种微阵列/100，000种化合物筛选）将用单独的酶和匹配的荧光底物激活。使用722个成员的荧光底物文库（Ala-P3-P2-K/R文库），在微阵列上对总共9种蛋白酶的底物特异性进行了充分分析。多重微阵列HTS的建议靶标是：丝氨酸蛋白酶（凝血）人因子IXa、Xla、Xa;丝氨酸蛋白酶（补体）人C1 s、C1 r、因子D;和人组织蛋白酶（溶酶体）组织蛋白酶B、S、G。在微阵列上使用多重测定允许产生化学蛋白酶相互作用组的关键数据。

DESCRIPTION (provided by applicant): A multiplexed microarray HTS campaign is proposed for a series of proteins relevant to blood coagulation, complement function, neutrophil platelet crosstalk, and lysosomal processes. Protocols are defined where the compound library will be printed in replicate sets on microarrays. Each set of microarrays (3072 compounds/microarray and 33 microarrays per 100,000 compound screen) will be activated with an individual enzyme and a matched fluorogenic substrate. A total of 9 proteases have been fully profiled on microarrays for substrate specificity using 722-member fluorogenic substrate libraries (Ala-P3-P2-K/R libraries). The proposed targets for multiplexed microarray HTS are: Serine proteases (Coagulation) Human factors IXa, Xla, Xa; Serine proteases (Complement) Human C1s, C1r, Factor D; and Human cathepsins (Lysosomal) Cathepsins B, S, G. The use of multiplexed assay on microarrays allows the generation of critical data for the chemical protease interactome.