Multiplexed Microarray Detection of Pluripotency Related Proteins

多能性相关蛋白的多重微阵列检测

• 批准号: 7610858
• 负责人: Scott Allen Monsma
• 金额: $ 18.46万
• 依托单位: PRIMORIGEN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7610858
• 关键词: Address; Affinity; Affinity Chromatography; Amino Acid Sequence; Antibodies; Binding; Biological Assay; Buffers; Caliber; Cell Extracts; Cells; Chemiluminescence assay; Data; Data Storage and Retrieval; Deposition; Detection; Development; Devices; Doctor of Philosophy; Drug Formulations; Ectoderm; Endoderm; Equilibrium; Evaluation; Figs - dietary; Funding; Government; Human; Human Cell Line; Immunoassay; Insecta; International; Ion-Exchange Chromatography Procedure; Ions; Kinetics; Letters; Life; Mammalian Cell; Measures; Mesoderm; Metals; Methods; Monoclonal Antibodies; Noise; Nuclear Extract; Oxidoreductase; Pathway Analysis; Peptide Sequence Determination; Performance; Persons; Phase; Phase I Clinical Trials; Plastics; Principal Investigator; Protein Array; Protein Binding; Proteins; Pyroxylin; Reagent; Recombinant Proteins; Recombinants; Reference Standards; Regulation; Research; Research Personnel; Sampling; Screening procedure; Small Business Innovation Research Grant; Specificity; Speed; Spottings; Stem cells; Surface; System; Technology; Testing; Thick; Time; Variant; assay development; base; beta Tubulin; cost; counterscreen; density; design; flexibility; gene cloning; high throughput screening; immunoaffinity chromatography; improved; milligram; miniaturize; novel; novel marker; pluripotency; pressure; public health relevance; research study; self-renewal

DESCRIPTION (provided by applicant): Primorigen Biosciences will use SBIR funds to develop a frameless, multiplexed, microarray screening system for rapid identification and characterization of antibody matched pairs with appropriate affinity and specificity for human proteins central to pluripotency and multipotency. This system produces improved assay content and dramatically reduces the time and cost of multiplexed microarray development. The core of this new system is Primorigen's novel frameless microarray device that allows multiplexed analysis of 96 (or 384) samples against several binding targets directly on a flat array surface. As a result, dozens of proteins in hundreds of samples can be detected with high precision to generate thousands of data points per experiment, increasing speed of screening. While the new system can be adapted for detection of any protein, this Phase I SBIR project will focus on characterizing assay content for detecting eight human cell markers of pluripotency and multipotency and validating them in a multiplexed array format. Phase II will a) complete additional microarrays of up to 15-20 multiplex assays, b) further develop automated methods to increase assay throughput, c) apply this system to pathway analysis involved in determining or maintaining cell potency and d) expand assay development to endoderm, mesoderm, and ectoderm markers. PUBLIC HEALTH RELEVANCE: Proteins facilitate many of the cell's most basic functions and thousands of new protein sequences and post-translational variants have been identified in the past few years. This protein jackpot has hastened the drive to understand protein-based regulation, but researchers require better biophysical methods to quantitatively detect protein markers. Primorigen's inexpensive, miniaturized, multiplexed, high throughput assays proposed for SBIR support will address this need, generating thousands of protein detection data points per experiment. At the same time, the project will provide new antibody and protein content for assays to detect human proteins central to pluripotency and multipotency.

描述（由申请人提供）： Primorigen Biosciences 将利用 SBIR 资金开发无框架、多重微阵列筛选系统，用于快速鉴定和表征抗体匹配对，对多能性和多功能性至关重要的人类蛋白质具有适当的亲和力和特异性。该系统可产生改进的检测内容，并显着减少多重微阵列开发的时间和成本。这个新系统的核心是 Primorigen 的新型无框微阵列装置，该装置允许直接在平面阵列表面上针对多个结合目标对 96 个（或 384 个）样本进行多重分析。因此，可以高精度检测数百个样品中的数十种蛋白质，从而在每个实验中生成数千个数据点，从而提高筛选速度。虽然新系统可以适用于检测任何蛋白质，但该 I 期 SBIR 项目将重点关注检测八种多能性和多能性人类细胞标记物的测定内容，并以多重阵列格式对其进行验证。 II 期将 a) 完成多达 15-20 次多重检测的额外微阵列，b) 进一步开发自动化方法以提高检测通量，c) 将该系统应用于涉及确定或维持细胞效力的途径分析，以及 d) 将检测开发扩展到内胚层、中胚层和外胚层标记。公共健康相关性：蛋白质促进细胞的许多最基本功能，并且在过去几年中已鉴定出数千种新蛋白质序列和翻译后变体。这种蛋白质大奖加速了理解基于蛋白质的调控的动力，但研究人员需要更好的生物物理方法来定量检测蛋白质标记。 Primorigen 为 SBIR 支持而提出的廉价、小型化、多重、高通量检测将满足这一需求，每个实验生成数千个蛋白质检测数据点。同时，该项目还将提供新的抗体和蛋白质含量，用于检测对多能性和多能性至关重要的人类蛋白质。