Electromobility Focusing for Separation of Proteins

描述（申请人提供）：血液中的蛋白质分布可以指示人体的总体状态或状况。 因此，血液样品中蛋白质的测定可以作为早期检测疾病状态（例如癌症）的有力诊断工具。不幸的是，目前还没有分析方法可以提供复杂混合物如血液中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迁移率聚焦（EMF）的新方法，用于以无与伦比的分辨率分离，浓缩和鉴定复杂混合物中的蛋白质。用于EMF的装置的物理描述是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，除了沿着柱的电场强度不是线性的，而是连续梯度。通过施加与蛋白质的电泳迁移方向相反的压力诱导液体流，可以在通道中以非常高的分辨率分离和聚焦蛋白质。作用在蛋白质上的合力将它们集中在沿着通道沿着的特定位置处的窄带中，以它们的电泳迁移率为顺序。通过改变压力诱导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设备格式进行EMF：（a）半制备型和（B）微芯片。 EMF可用于血清中前列腺特异性抗原、癌胚抗原、癌相关抗原1 - 25、甲胎蛋白和人绒毛膜促性腺激素等5种已知蛋白质肿瘤标志物的快速定量检测。还将研究拟议的新肿瘤标志物胸苷激酶I。

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.