An Automated 2D-Capillary Electrophoresis System

自动化二维毛细管电泳系统

• 批准号: 6480211
• 负责人: GARY LOGE
• 金额: $ 14.98万
• 依托单位: SPECTRUMEDIX CORPORATION
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6480211
• 关键词: bioengineering /biomedical engineering; biomedical automation; biomedical equipment development; capillary electrophoresis; electrofocusing; fluorescence; proteomics

DESCRIPTION (provided by applicant): For complex protein mixtures, including cell lysates, containing thousands of proteins, 2-D gel electrophoresis separation (2D-PAGE) is the standard method used to separate and isolate individual proteins for analysis. In addition to being useful for identifying individual proteins, it is also used to monitor changes in the amount of a specific protein or proteins as an indication of the onset of cancer in tissue or how cancer growth is affected by environmental influences or by treatment methods. We will develop a capillary equivalent to the standard 2D-PAGE method. It will offer several advantages compared to 2 D-PAGE, including automation, quantitative measurement in individual proteins, dramatically decreased run time, and reproducible size and pI measurement for protein identification. The main innovation is a scheme that will couple capillary iso-electric focusing (CIEF) with multicapillary capillary-gel electrophoresis (CGE) to allow automated two-dimensional capillary-gel electrophoresis of protein samples to be performed. In Phase I, the coupling scheme will be developed and tested. In addition, the fluorescence detection chemistry using "on-column" detection on the capillary array will be developed. This will be a significant advance for the analysis of whole cell samples for clinical and research studies of cancerous and pre-cancerous tissue. PROPOSED COMMERCIAL APPLICATION: Many clinical and research labs involved in cancer research as well as proteomics research would benefit from the type of instrument that we are proposing to develop. Clinical labs for routine analysis would be our primary target, although a second design with more user control and flexibility for research applications would probably be the initial market.

描述（由申请人提供）： 对于复杂的蛋白质混合物，包括细胞裂解物，含有数千个 蛋白质，2-D凝胶电泳分离（2D-PAGE）是标准方法 用于分离和分离单个蛋白质以进行分析。除了 它可用于识别单个蛋白质，也可用于监测 特定蛋白质或蛋白质的量的变化作为 组织中癌症的发生或癌症生长如何受到 环境影响或处理方法。 我们将开发一种与标准2D-PAGE方法相当的毛细管。它将 与2D-PAGE相比，提供了几个优点，包括自动化， 单个蛋白质的定量测量， 用于蛋白质鉴定的可重复的大小和pI测量。的 主要创新是一个方案，将耦合毛细管等电聚焦 （CIEF）与多毛细管毛细管凝胶电泳（CGE），以允许 蛋白质样品的自动化二维毛细管凝胶电泳， 被执行。在第一阶段，将制定和测试耦合方案。在 此外，使用“柱上”检测的荧光检测化学 将开发毛细管阵列。 这将是分析全细胞样品的重大进展， 癌症和癌前组织的临床和研究。 拟定商业应用：许多参与癌症研究和蛋白质组学研究的临床和研究实验室将受益于我们提议开发的仪器类型。 用于常规分析的临床实验室将是我们的主要目标，尽管具有更多用户控制和研究应用灵活性的第二种设计可能是最初的市场。

项目成果

Poly(2-ethyl-2-oxazoline) as a sieving matrix for SDS-CE.

• DOI: 10.1002/elps.200900107
• 发表时间: 2009-12
• 期刊: ELECTROPHORESIS
• 影响因子: 2.9
• 作者: Bernard, Richard;Loge, Gary
• 通讯作者: Loge, Gary