Electromobility Focusing for Separation of Proteins

用于蛋白质分离的电动聚焦

• 批准号: 6849275
• 负责人: MILTON L. LEE
• 金额: $ 33.93万
• 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/6849275
• 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（由申请人提供）：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疾病状态（例如癌症）的强大诊断工具。不幸的是，当今尚无分析方法可提供复杂混合物（如血液）中存在的蛋白质的快速高分辨率曲线。该提案的总体目的是开发一种新方法，称为电动性聚焦（EMF），用于分离，浓缩和鉴定具有无与伦比分辨率的复杂混合物中的蛋白质。 EMF设备的物理描述是一个通道，其中包含沿着高压的缓冲溶液，类似于毛细管电泳，只是沿柱沿线的电场强度不是线性的，而是连续的梯度。通过将压力诱导的液体流动计数器应用于蛋白质的电泳迁移方向，可以将蛋白质分离并聚焦在通道中的高分辨率。作用在蛋白质上的作用力将它们集中在沿通道的特定位置的狭窄带中，以其电泳迁移率。可以通过更改压力诱导的反流的大小或通过降低施加的电压来动员蛋白质。这项工作的特定目标包括开发用于执行EMF的两种不同的设备格式：（a）半预编程和（b）微芯片。 EMF将应用于血清中五种已知蛋白肿瘤标记物的快速定量检测，包括前列腺特异性抗原，癌胚抗原，与癌相关的抗原1 25，α-五蛋白蛋白和雄鹿科氏蛋白原。还将研究拟议的新肿瘤标记胸苷激酶I。