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    • 8. 发明申请
    • STERILITY INDICATING BIOLOGICAL COMPOSITIONS, ARTICLES AND METHODS
    • 表明生物组成的无效性,文章和方法
    • US20110195442A1
    • 2011-08-11
    • US13123824
    • 2009-10-15
    • Sailaja ChandrapatiHeather M. WebbAi-Ping Wei
    • Sailaja ChandrapatiHeather M. WebbAi-Ping Wei
    • C12Q1/37
    • C12Q1/37A61L2/28C12Q1/22
    • A sterility indicating composition comprising a plurality of sterilization process resistant spores which contain an active protease during germination and initial outgrowth of the spores; and a germination medium comprising at least one labeled protease substrate and at least one nutrient for germination of the spores; wherein the medium is essentially free of a) any active protease other than the active protease contained by the plurality of spores and b) any protease substrate other than the at least one labeled protease substrate, other than any protease substrate originating from the plurality of spores, and other than any protease substrate which does not compete with the labeled protease substrate for the active protease; and wherein the at least one labeled protease substrate comprises a peptide which can be cleaved by the active protease and which is labeled with one or more dye groups, at least one of which undergoes a detectable change when the peptide is cleaved by the active protease, and wherein the labeled protease substrate is stable at least at a temperature for incubating the spores, a sterilization process indicator comprising the composition, and a method of determining the effectiveness of a sterilization process using the composition and indicator are disclosed.
    • 一种不育指示组合物,其包含多个灭菌过程抗性孢子,其在萌发期间含有活性蛋白酶和孢子的初始产生; 以及包含至少一种标记的蛋白酶底物和至少一种用于孢子发芽的营养物的发芽培养基; 其中所述培养基基本上不含a)除所述多个孢子所含的活性蛋白酶以外的任何活性蛋白酶,和b)除所述至少一个标记的蛋白酶底物之外的任何蛋白酶底物,而不是源于所述多个孢子的任何蛋白酶底物 ,而不是与活性蛋白酶的标记的蛋白酶底物竞争的任何蛋白酶底物以外的其它蛋白酶底物; 并且其中所述至少一个标记的蛋白酶底物包含可被活性蛋白酶切割并且被一个或多个染料基团标记的肽,当肽被活性蛋白酶切割时,其至少一种经历可检测的变化, 并且其中标记的蛋白酶底物至少在用于培养孢子的温度下是稳定的,包括该组合物的灭菌过程指示剂以及使用该组合物和指示剂确定灭菌过程的有效性的方法。
    • 10. 发明授权
    • Fluorogenic protease substrates based on dye-dimerization
    • 基于染料二聚化的荧光蛋白酶底物
    • US07256012B2
    • 2007-08-14
    • US10930522
    • 2004-08-31
    • Ai-Ping WeiMichael George Williams
    • Ai-Ping WeiMichael George Williams
    • C12Q1/37C12Q1/04C07K7/06G01N2/64
    • C07K7/06C12Q1/37C12Q2337/40Y02A50/451
    • A method of biological assay comprises the steps of providing an enzyme substrate comprising two fluorescence dye groups bound to a flexible peptide, the dye groups being of proximity sufficiently close so as to allow free energy attractions to draw the dyes together to essentially self-quench fluorescence of the dye groups, wherein self quenching of fluorescence of the dye groups is effected by dye dimerization or stacking, and enzymatically cleaving the peptide to release the fluorescence dye groups from dye dimerization or stacking, thereby producing an increase in fluorescence intensity. A protease substrate for use in the method of the invention is also disclosed. This invention finds use in detection and identification of microorganisms, sterilization assurance, pharmaceutical discovery, enzyme assays, immunoassays, and other biological assays.
    • 一种生物测定方法包括以下步骤:提供包含与柔性肽结合的两个荧光染料基团的酶底物,所述染料组的接近度足够接近,以便使自由能吸引力将染料一起吸收到基本上自熄灭荧光 的染料基团,其中染料基团的荧光自发淬灭通过染料二聚化或堆叠进行,并且酶切割肽以使荧光染料基团从染料二聚化或堆叠释放,从而产生荧光强度的增加。 还公开了用于本发明方法的蛋白酶底物。 本发明用于微生物的检测和鉴定,灭菌保证,药物发现,酶测定,免疫测定和其他生物测定。