Development of a novel catalytic array

新型催化阵列的开发

• 批准号: 7325833
• 负责人: Tauseef R. Butt
• 金额: $ 26.38万
• 依托单位: LIFESENSORS, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-20 至 2009-09-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7325833
• 关键词: Antibodies; Antibody Formation; Autoantibodies; Basic Science; Binding; Bioinformatics; Biological Assay; Biological Markers; Cell Survival; Class; Complex; Condition; Development; Devices; Diabetes Mellitus; Diagnosis; Diagnostic; Disease; Enhancement Technology; Enzymes; General Population; Genomics; Goals; Homeostasis; Immobilization; Malignant Neoplasms; Methods; Numbers; Parkinsonian Disorders; Pathway interactions; Phase; Play; Protein Microchips; Proteins; Purpose; Reagent; Relative (related person); Research Personnel; Role; Sampling; Screening procedure; Serum; Solid; Staging; Standards of Weights and Measures; System; Therapeutic; Ubiquitin; Ubiquitin Like Proteins; Validation; Variant; cost; cost effective; enzyme activity; high throughput screening; inhibitor/antagonist; interest; novel; outcome forecast; prognostic; protein expression; therapeutic target; tool; ubiquitin isopeptidase

DESCRIPTION (provided by applicant): Bioinformatics tools will be generated for a class of ~ 100 proteins known as de- ubiquitinating enzymes (DUBs), which play an important role in cell survival, proliferation, and differentiation. LifeSensor's proprietary SUMO expression enhancement technology permits expression and purification of DUBs with relative ease, and a high throughput microarray of these proteins will afford rapid and cost effective screening. Thus, it is proposed to develop a DUB microarray for the purpose of basic research, biomarker discovery, diagnostics/prognostics, and therapeutics. The DUB microarray will have obvious utility in biomarker discovery and diagnostics development. Expression of DUBs has been associated with diseases such as cancer, diabetes, and Parkinsonism. Over- or under-expression of proteins in disease states tend to elicit elevated or diminished auto-antibody production against those proteins. Thus, auto-antibody profiling can assist in identifying the type and stage of a disease. When a biomarker is identified, it can be further studied for its role in the diagnosis and prognosis of the disease. The particular biomarker could also be a therapeutic target. A DUB microarry not only allows one to perform high throughput screening of samples, but also to evaluate expression levels of DUBs in samples under identical conditions to correlate their effect on each other's expression levels under different circumstances. In the first aim, 10 (ten) DUBs will be immobilized on a variety of solid supports to determine the optimum support in terms of retention of enzyme activity. Next, the catalytic function of the retained DUBs will be compared with their function in a standard assay system. Finally, 50 (fifty) serum samples from the general population will be profiled for variation in the expression of autoantibodies against the DUBs. In Phase II, arrays will be built consisting of all known DUBS for ubiquitin and relevant ubiquitin-like proteins.

描述（由申请人提供）：将生成生物信息学工具，用于〜100种蛋白质的类别，称为泛素化酶（DUBS），这些酶在细胞存活，增殖和分化中起着重要作用。 Lifesensor的专有SUMO表达增强技术允许相对轻松地表达和纯化配音，并且这些蛋白质的高吞吐量微阵列将提供快速且具有成本效益的筛选。因此，建议开发一个配音微阵列，以实现基础研究，生物标志物发现，诊断/预后和治疗学的目的。配音微阵列将在生物标志物发现和诊断开发方面具有明显的实用性。 DUB的表达与癌症，糖尿病和帕金森氏症等疾病有关。疾病状态中蛋白质的过度表达往往会引起针对这些蛋白质的自身抗体产生升高或减少。因此，自身抗体分析可以帮助识别疾病的类型和阶段。当发现生物标志物时，可以进一步研究其在疾病的诊断和预后中的作用。特定的生物标志物也可能是治疗靶标。 DUB微型携带不仅允许一个人对样品进行高吞吐量筛选，还可以评估样品在相同条件下样品中的配音水平，以将它们在不同情况下对彼此表达水平的影响相关联。在第一个目标中，将在各种固体支持上固定10（十）个配音，以确定保留酶活性的最佳支持。接下来，将保留配音的催化功能与标准测定系统中的功能进行比较。最后，将介绍来自一般人群的50个（五十）血清样品，以构建自身抗体对DUB的表达的变化。在第二阶段，将建立由所有已知的泛素和相关泛素样蛋白组成的阵列。