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.

血液中的蛋白质分布可以指示人体的整体状态或状况。 因此，测定血液样品中的蛋白质可以作为早期糖尿病的强有力的诊断工具。 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 以提供复杂混合物如血液中存在的蛋白质的快速高分辨率图谱。整体 该建议的目的是开发一种称为电迁移率聚焦（EMF）的新方法， 浓缩，并以无与伦比的分辨率鉴定复杂混合物中的蛋白质。物理 用于EMF的装置的描述是含有缓冲溶液的通道，高电压沿着通道被施加。 类似于毛细管电泳，除了沿着柱沿着的电场强度不 线性的，但是是连续的梯度。蛋白质可以分离和聚焦， 通过将压力诱导的液体流计数器应用于电泳， 蛋白质的迁移方向。作用在蛋白质上的合力将它们集中成 在沿着沟道的特定位置处的窄带，其顺序为它们的电泳迁移率。的 蛋白质可以通过改变压力诱导的流动的大小而在分离通道中移动 反向流动或通过降低所施加的电压。这项工作的具体目标包括： 开发了两种不同的用于执行EMF的装置形式：（a）半制备型和（B）微芯片。 将EMF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、α- 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， 也被研究。

项目成果

Field gradient electrophoresis.

场梯度电泳。

• DOI: 10.1002/elps.200410319
• 发表时间: 2005
• 期刊: Electrophoresis.
• 作者: Warnick,KarlF;Francom,ScottJ;Humble,PaulH;Kelly,RyanT;Woolley,AdamT;Lee,MiltonL;Tolley,HDennis
• 通讯作者: Tolley,HDennis

Performance optimization in electric field gradient focusing.

电场梯度聚焦的性能优化。

• DOI: 10.1016/j.chroma.2008.11.031
• 发表时间: 2009
• 期刊: Journal of chromatography. A
• 作者: Sun,Xuefei;Farnsworth,PaulB;Tolley,HDennis;Warnick,KarlF;Woolley,AdamT;Lee,MiltonL
• 通讯作者: Lee,MiltonL

Poly(ethylene glycol)-functionalized polymeric microchips for capillary electrophoresis.

• DOI: 10.1021/ac9001832
• 发表时间: 2009-07
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Xuefei Sun;Dan-dan Li;Milton L. Lee

Surface-reactive acrylic copolymer for fabrication of microfluidic devices.

用于制造微流体装置的表面反应性丙烯酸共聚物。

• DOI: 10.1021/ac0580060
• 发表时间: 2005
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Liu,Jikun;Sun,Xuefei;Lee,MiltonL
• 通讯作者: Lee,MiltonL

Surface modification of glycidyl-containing poly(methyl methacrylate) microchips using surface-initiated atom-transfer radical polymerization.

使用表面引发的原子转移自由基聚合对含缩水甘油基的聚（甲基丙烯酸甲酯）微芯片进行表面改性。

• DOI: 10.1021/ac701948n
• 发表时间: 2008
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Sun,Xuefei;Liu,Jikun;Lee,MiltonL
• 通讯作者: Lee,MiltonL