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    • 4. 发明申请
    • Methods for conducting assays for enzyme activity on protein microarrays
    • 蛋白质微阵列酶活性测定方法
    • US20050118665A1
    • 2005-06-02
    • US10865431
    • 2004-06-09
    • Fang ZhouBarry Schweitzer
    • Fang ZhouBarry Schweitzer
    • C12Q1/00C12Q1/68C12Q1/37
    • C12Q1/00
    • The present invention relates to methods of conducting assays for enzymatic activity on microarrays useful for the large-scale study of protein function, screening assays, and high-throughput analysis of enzymatic reactions. The invention relates to methods of using protein chips to assay the presence, amount, activity and/or function of enzymes present in a protein sample on a protein chip. In particular, the methods of the invention relate to conducting enzymatic assays using a microarray wherein a protein and a substance are immobilized on the surface of a solid support and wherein the protein and the substance are in proximity to each other sufficient for the occurrence of an enzymatic reaction between the substance and the protein. The invention also relates to microarrays that have an enzyme and a substrate immobilized on their surface wherein the enzyme and the substrate are in proximity to each other sufficient for the occurrence of an enzymatic reaction between the enzyme and the substrate.
    • 本发明涉及对用于蛋白质功能的大规模研究,筛选测定和酶促反应的高通量分析的微阵列上的酶活性进行测定的方法。 本发明涉及使用蛋白质芯片来测定存在于蛋白质芯片上的蛋白质样品中的酶的存在,量,活性和/或功能的方法。 特别地,本发明的方法涉及使用微阵列进行酶测定,其中将蛋白质和物质固定在固体支持物的表面上,并且其中所述蛋白质和所述物质彼此接近,足以发生 物质与蛋白质之间的酶反应。 本发明还涉及具有固定在其表面上的酶和底物的微阵列,其中酶和底物相互接近,足以在酶和底物之间发生酶反应。
    • 8. 发明申请
    • Protein expression profiling
    • 蛋白质表达谱
    • US20060166227A1
    • 2006-07-27
    • US11187537
    • 2005-07-22
    • Stephen KingsmoreGirish NallurBarry Schweitzer
    • Stephen KingsmoreGirish NallurBarry Schweitzer
    • C12Q1/68C12P19/34
    • C12Q1/6816C12Q1/6837C12Q1/6844C12Q2565/501C12Q2531/125
    • Disclosed are compositions and methods for detecting small quantities of analytes such as proteins and peptides. The method involves associating a primer with an analyte and subsequently using the primer to mediate rolling circle replication of a circular DNA molecule. Amplification of the DNA circle is dependent on the presence of the primer. Thus, the disclosed method produces an amplified signal, via rolling circle amplification, from any analyte of interest. The amplified DNA remains associated with the analyte, via the primer, and so allows spatial detection of the analyte. The disclosed method can be used to detect and analyze proteins and peptides. Multiple proteins can be analyzed using microarrays to which the various proteins are immobilized. A rolling circle replication primer is then associated with the various proteins using a conjugate of the primer and a molecule that specifically binds the proteins to be detectable. Rolling circle replication from the primers results in production of a large amount of DNA at the sites in the array where the proteins are immobilized. The DNA produced by rolling circle replication can be further amplified in secondary and higher order amplification processes using second-stage or higher order primers in conjunction with second-stage or higher order amplification target circles. The amplified DNA serves as a readily detectable signal for the proteins. The disclosed method can also be used to compare the proteins expressed in two or more different samples. The information generated is analogous to the type of information gathered in nucleic acid expression profiles. The disclosed method allows sensitive and accurate detection and quantitation of proteins expressed in any cell or tissue.
    • 公开了用于检测少量分析物如蛋白质和肽的组合物和方法。 该方法包括将底物与分析物缔合,随后使用引物介导环状DNA分子的滚环复制。 DNA环的扩增取决于引物的存在。 因此,所公开的方法从感兴趣的任何分析物通过循环扩增产生扩增信号。 扩增的DNA通过引物与分析物保持相关,因此允许分析物的空间检测。 所公开的方法可用于检测和分析蛋白质和肽。 可以使用固定有各种蛋白质的微阵列来分析多种蛋白质。 然后使用引物和特异性结合蛋白质的可检测的分子的缀合物将滚环复制引物与各种蛋白质相关联。 来自引物的滚环复制导致在固定有蛋白质的阵列中的位点产生大量的DNA。 通过循环复制产生的DNA可以在次级和高级扩增过程中进一步扩增,使用第二级或更高级引物连同第二级或更高级扩增目标圆。 扩增的DNA作为蛋白质的易检测信号。 所公开的方法也可用于比较在两个或多个不同样品中表达的蛋白质。 产生的信息类似于在核酸表达谱中收集的信息的类型。 所公开的方法允许敏感且准确地检测和定量在任何细胞或组织中表达的蛋白质。