Electromobility Focusing for Separation of Proteins

用于蛋白质分离的电动聚焦

• 批准号: 6698842
• 负责人: MILTON L. LEE
• 金额: $ 36.89万
• 依托单位: BRIGHAM YOUNG UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6698842
• 关键词: alpha fetoprotein; biomarker; blood chemistry; blood proteins; carcinoembryonal antigen; chorionic gonadotropin; clinical research; electrofocusing; human tissue; prostate specific antigen; protein purification; serology /serodiagnosis; technology /technique development; thymidine kinase

DESCRIPTION (provided by applicant): The protein distribution in the blood can indicate the overall state or condition of the human body. Therefore, determination of the proteins in blood samples can serve as a powerful diagnostic tool for early detection of disease states, such as cancer. Unfortunately, there is no analytical method available today to provide rapid high resolution profiles of proteins present in complex mixtures such as blood. The overall objective of this proposal is to develop a new method called electromobility focusing (EMF) for separating, concentrating, and identifying proteins in complex mixtures with unsurpassed resolution. The physical description of the device for EMF is a channel containing a buffer solution along which a high voltage is applied, similar to capillary electrophoresis, except that the electric field intensity along the column is not linear, but is a continuous gradient. Proteins can be separated and focused with exceptionally high resolution in the channel by applying a pressure-induced liquid flow counter to the electrophoretic migration direction of the proteins. The resulting forces acting on the proteins concentrate them into narrow bands at specific positions along the channel in order of their electrophoretic mobilities. The proteins can be mobilized in the separation channel by changing the magnitude of the pressure-induced counter flow or by reducing the applied voltage. The specific objectives of this work include the development of two different device formats for performing EMF: (a) semi-preparative and (b) microchip. EMF will be applied to the rapid quantitative detection of five known protein tumor markers in blood serum, including prostate specific antigen, carcinoembryonic antigen, carcinoma-associated antigen 1 25, alpha-fetoprotein, and humanchorionic gonadotropin. Proposed new tumor marker, thymidine kinase I, will also be studied.

描述（申请人提供）：血液中的蛋白质分布可以表明人体的整体状态或状况。 因此，血液样本中蛋白质的测定可以作为早期检测癌症等疾病状态的强大诊断工具。不幸的是，目前还没有可用的分析方法来提供复杂混合物（例如血液）中存在的蛋白质的快速高分辨率图谱。该提案的总体目标是开发一种称为电动聚焦 (EMF) 的新方法，用于以无与伦比的分辨率分离、浓缩和识别复杂混合物中的蛋白质。 EMF装置的物理描述是一个含有缓冲溶液的通道，沿着该通道施加高电压，类似于毛细管电泳，不同之处在于沿柱的电场强度不是线性的，而是连续梯度的。通过施加与蛋白质电泳迁移方向相反的压力诱导液流，可以在通道中以极高的分辨率分离和聚焦蛋白质。作用在蛋白质上的力将它们按照电泳迁移率的顺序集中在通道上特定位置的窄带中。通过改变压力引起的逆流的大小或通过降低施加的电压，可以在分离通道中移动蛋白质。这项工作的具体目标包括开发两种不同的用于执行 EMF 的设备格式：(a) 半制备型和 (b) 微芯片。 EMF将应用于血清中5种已知蛋白肿瘤标志物的快速定量检测，包括前列腺特异性抗原、癌胚抗原、癌相关抗原1 25、甲胎蛋白、人绒毛膜促性腺激素。拟议的新肿瘤标志物胸苷激酶 I 也将被研究。