Electromobility Focusing for Separation of Proteins

血液中蛋白质的分布可以指示人体的整体状态或状况。 因此，血样中蛋白质的测定可作为早期诊断的有力工具。 检测疾病状态，例如癌症。不幸的是，目前还没有可用的分析方法。 以提供血液等复杂混合物中存在的蛋白质的快速高分辨率图谱。整体而言 该方案的目的是开发一种新的方法，称为电迁移率聚焦(EMF)，用于分离， 以无与伦比的分辨率浓缩和识别复杂混合物中的蛋白质。体能 用于电磁场的装置的描述是包含缓冲溶液的通道，沿该缓冲溶液高电压 应用，类似于毛细管电泳法，只是沿柱的电场强度不是 线性，但是是一个连续的渐变。蛋白质可以被分离并以极高的浓度集中 通过将压力诱导液流计数器应用于电泳来实现通道中的分辨率 蛋白质的迁移方向。作用在蛋白质上的作用力将它们浓缩成 沿通道的特定位置上的窄带，按其电泳迁移率的顺序排列。这个 通过改变压力诱导的大小，可以在分离通道中动员蛋白质 逆流或通过降低施加的电压。这项工作的具体目标包括 开发两种不同的执行电磁场的设备格式：(A)半制备和(B)微芯片。 电磁场将应用于血清中五种已知蛋白质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125、 胎儿蛋白和人绒毛膜促性腺激素。一种新的肿瘤标记物，胸苷激酶I，将 也有待研究。

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. Aproposed new tumor marker, thymidine kinase I, will also be studied.