Bioanalytical Applications of Capillary Electrophoresis

毛细管电泳的生物分析应用

• 批准号: 6953816
• 负责人: FRANK A GOMEZ
• 金额: $ 14.12万
• 依托单位: CALIFORNIA STATE UNIVERSITY LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6953816
• 关键词: analytical method; capillary electrophoresis; chemical kinetics; chemical property; high performance liquid chromatography; high throughput technology; method development; physical property; ristocetin; small molecule; vancomycin

DESCRIPTION (provided by applicant): The broad goals of this research are to further develop capillary electrophoresis (CE) as an analytical technique to study the physicochemical properties of the glycopeptide antibiotics vancomycin (Van), ristocetin (Ris), and teicoplanin (Tei) and their derivatives. Its focus is on demonstrating principles and developing useful analytical techniques that can be applied to the analysis of receptor-ligand interactions and microscale reactions. The development of resistance to antibacterial agents is a worldwide problem that continues to compromise the clinical effectiveness of new drugs used in the treatment of many infectious diseases. Hence, studies probing the physicochemical properties of these antibiotics via chemical modification are needed. In the proposed research two specific CE techniques will be: on-column microreactor techniques and affinity capillary electrophoresis (ACE). Techniques utilized in this study include chemical modification and characterization, chemical separation and identification of small molecules by CE utilizing both laser-induced fluorescence (LIF) and ultraviolet/visible (UV/VIS) detection schemes and high-performance liquid chromatography (HPLC). The research will develop new bioanalytical techniques and will focus on examining small biomolecules involved in the prevention of disease and, hence, to the issue of public health. This understanding will strengthen the scientific base underlying the design, preparation, and application of CE towards a host of health-related problems. The specific aims of the research are to I. Couple On-Column Linked Antibiotic Derivatization Reactions to ACE. II. Examine On-Column Ligand Derivatization Reactions Coupled to ACE. III. Utilize Multiple-Injection ACE (MIACE) to Analyze the Binding of Ligands to Glycopeptide Antibiotics. IV. Optimize Conditions for Flow-Through Partial-Filling ACE (FTPFACE) to Analyze the Binding of Ligands to Antibiotics. The proposed research will: Demonstrate the versatility of CE in determining physicochemical parameters of antibiotics; Demonstrate high-throughput derivatization of receptors and ligarids coupled to ACE; Provide for rapid synthesis and accurate analysis of drug targets; require reduced samples volumes compared to other analytical techniques and; reduce sample waste and disposal.

描述（由申请方提供）：本研究的广泛目标是进一步开发毛细管电泳（CE）作为一种分析技术，以研究糖肽类抗生素万古霉素（货车）、瑞斯托霉素（Ris）和替考拉宁（Tei）及其衍生物的理化性质。它的重点是展示原理和开发有用的分析技术，可应用于受体-配体相互作用和微尺度反应的分析。对抗菌剂的耐药性的发展是一个世界性的问题，其继续损害用于治疗许多感染性疾病的新药的临床有效性。因此，需要通过化学修饰来探索这些抗生素的理化性质的研究。在拟议的研究中，两个具体的CE技术将是：柱上微反应器技术和亲和毛细管电泳（ACE）。本研究中使用的技术包括化学修饰和表征、通过CE利用激光诱导荧光（LIF）和紫外/可见光（UV/维斯）检测方案以及高效液相色谱法（HPLC）进行小分子的化学分离和鉴定。该研究将开发新的生物分析技术，并将重点放在检查参与预防疾病的小生物分子，从而解决公共卫生问题。这种理解将加强CE设计、制备和应用于许多健康相关问题的科学基础。本研究的具体目的是：偶联柱上连接的抗生素衍生化反应至ACE。二.检查与ACE偶联的柱上配体衍生化反应。三.利用多次注射ACE（MIACE）分析配体与糖肽类抗生素的结合。四.优化用于分析配体与抗生素结合的流通部分填充ACE（FTPFACE）的条件。拟议的研究将：证明CE在确定抗生素理化参数方面的多功能性;证明与ACE偶联的受体和Ligarids的高通量衍生化;提供药物靶标的快速合成和准确分析;与其他分析技术相比，需要减少的样品体积;减少样品浪费和处置。

项目成果

Fabrication of fritless chromatographic microchips packed with conventional reversed-phase silica particles.

制造填充有传统反相二氧化硅颗粒的无熔块色谱微芯片。

• DOI: 10.1021/ac071106g
• 发表时间: 2007
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Gaspar,Attila;Piyasena,MenakeE;Gomez,FrankA
• 通讯作者: Gomez,FrankA

Design and development of a flow injection-capillary electrophoresis analyzer employing fiber optic detection.

采用光纤检测的流动注射毛细管电泳分析仪的设计和开发。

• 发表时间: 2007
• 期刊: Journal of capillary electrophoresis and microchip technology
• 作者: Hanrahan,Grady;Tse,Florence;Dahdouh,FroseenT;Clarke,Keith;Gomez,FrankA
• 通讯作者: Gomez,FrankA

Use of voltage gradient partial-filling affinity capillary electrophoresis to estimate binding constants of ligands to receptors.

使用电压梯度部分填充亲和毛细管电泳来估计配体与受体的结合常数。

• 发表时间: 2007
• 期刊: Journal of capillary electrophoresis and microchip technology
• 作者: Ramirez,Alejandra;Gomez,FrankA
• 通讯作者: Gomez,FrankA

Magnetic microsphere-based methods to study the interaction of teicoplanin with peptides and bacteria.

• DOI: 10.1007/s00216-008-2327-y
• 发表时间: 2008-11
• 期刊: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
• 影响因子: 4.3
• 作者: Piyasena, Menake E.;Real, Lilian J.;Diamond, Rochelle A.;Xu, H. Howard;Gomez, Frank A.
• 通讯作者: Gomez, Frank A.

Through-a-chip partial filling affinity capillary electrophoresis for estimating binding constants of ligands to receptors.

通过芯片部分填充亲和毛细管电泳，用于估计配体与受体的结合常数。

• DOI: 10.1016/j.talanta.2007.06.043
• 发表时间: 2008
• 期刊: Talanta
• 影响因子: 6.1
• 作者: Brown,Abby;Morales,Christina;Gomez,FrankA
• 通讯作者: Gomez,FrankA