Electromobility Focusing for Separation of Proteins

用于蛋白质分离的电动聚焦

• 批准号: 7616933
• 负责人: MILTON L. LEE
• 金额: $ 11.67万
• 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 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7616933
• 关键词: AFP gene; Biological; Blood; Blood specimen; Buffers; Capillary Electrophoresis; Carcinoembryonic Antigen; Colorectal Cancer; Complex; Complex Mixtures; Condition; Detection; Development; Devices; Diagnostic; Disease; Early Diagnosis; Fetoprotein; Gonadotropins; Hepatic; Human Chorionic Gonadotropin; Human body; Liquid substance; Malignant Neoplasms; Malignant neoplasm of ovary; Malignant neoplasm of prostate; Malignant neoplasm of testis; Methods; Modeling; Operative Surgical Procedures; Performance; Positioning Attribute; Prostate-Specific Antigen; Protein Analysis; Proteins; Resolution; Sampling; Serum; Shapes; Solutions; Testing; Thymidine Kinase; Today; Tumor Markers; Tumor-Associated Carbohydrate Antigens; Work; alpha-Fetoproteins; analytical method; aqueous; electric field; malignant breast neoplasm; microchip; migration; pressure; protein distribution; tool; voltage

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.

血液中蛋白质的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诊断的有力诊断工具。 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 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。整体 该提案的目标是开发一种称为电动聚焦（EMF）的新方法，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。物理方面 EMF 设备的描述是一个包含缓冲溶液的通道，沿该通道施加高电压 应用，类似于毛细管电泳，不同之处在于沿柱的电场强度不 线性，但是是连续的梯度。可以以极高的速度分离和聚焦蛋白质 通过对电泳施加压力诱导液流计数器来提高通道中的分辨率 蛋白质的迁移方向。由此产生的作用在蛋白质上的力将它们集中到 沿着通道的特定位置处的窄带按照其电泳迁移率的顺序。这 通过改变压力诱导的大小，可以在分离通道中移动蛋白质 逆流或通过降低施加的电压。这项工作的具体目标包括 开发用于执行 EMF 的两种不同设备格式：(a) 半制备型和 (b) 微芯片。 EMF将应用于血清中五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、α- 甲胎蛋白和人绒毛膜促性腺激素。拟议的新肿瘤标志物胸苷激酶 I 将 也有待研究。

项目成果

Design and evaluation of a coupled monolithic pre-concentrator-capillary zone electrophoresis system for the extraction of immunoglobulin G from human serum.

用于从人血清中提取免疫球蛋白 G 的耦合整体式预浓缩器-毛细管区带电泳系统的设计和评估。

• 发表时间: 2005-12-02
• 作者: Armenta, Jenny M;Gu, Binghe;Humble, Paul H;Thulin, Craig D;Lee, Milton L
• 通讯作者: Lee, Milton L

Efficient polymer monolith for strong cation-exchange capillary liquid chromatography of peptides.

用于肽的强阳离子交换毛细管液相色谱的高效聚合物整体。

• 发表时间: 2006-06-01
• 影响因子: 7.4
• 作者: Gu, Binghe;Chen, Zhaoyuan;Thulin, Craig D;Lee, Milton L
• 通讯作者: Lee, Milton L

Field gradient electrophoresis.

场梯度电泳。

• 发表时间: 2005-01
• 作者: Warnick, Karl F;Francom, Scott J;Humble, Paul H;Kelly, Ryan T;Woolley, Adam T;Lee, Milton L;Tolley, H Dennis
• 通讯作者: Tolley, H Dennis

Electric field gradient focusing.

电场梯度聚焦。

• 发表时间: 2005-10
• 作者: Kelly, Ryan T;Woolley, Adam T
• 通讯作者: Woolley, Adam T

Performance optimization in electric field gradient focusing.

电场梯度聚焦的性能优化。

• 发表时间: 2009-01-02
• 作者: Sun, Xuefei;Farnsworth, Paul B;Tolley, H Dennis;Warnick, Karl F;Woolley, Adam T;Lee, Milton L
• 通讯作者: Lee, Milton L