Capillary Electrophoresis: Bioanalytical Applications

毛细管电泳：生物分析应用

• 批准号: 6668822
• 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: 2003
• 资助国家: 美国
• 起止时间: 2003-07-15 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6668822
• 关键词: analytical chemistry; antibiotics; capillary electrophoresis; chemical kinetics; chemical reaction; chemical synthesis; combinatorial chemistry; drug resistance; high performance liquid chromatography; high throughput technology; intermolecular interaction; ligands; receptor; ristocetin; technology /technique development; vancomycin

DESCRIPTION (provided by applicant): The broad goals of this research are to develop capillary electrophoresis (CE) as an analytical technique to study the physicochemical parameters of the antibiotics vancomycin (Van), ristocetin (Ris), and teicoplanin (Tel). Its focus is on demonstrating principles and on developing useful bioanalytical procedures that can be applied to the analysis of receptor-ligand interactions and microscale reactions. The evolution of antibiotic resistant bacteria has made the study and development of Van-like antibiotics a critical area of research focus. Hence, studies probing the physicochemical parameters of these antibiotics via chemical modification are highly warranted. Two specific CE techniques will be utilized in this work: 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), and enzyme kinetics. 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. Utilize On-Column Microreactor Techniques and ACE in the Estimation of Binding Constants Between Antibiotics and Ligands. II. Determine the Charge and pl of Antibiotics Using On-Column Microreactor Techniques. III. Examine the Kinetics of Derivatization of Antibiotics Using Reactive Chemical Reagents. IV. Estimate Binding Constants of Linked Antibiotics to Peptides. V. Conduct Experimental Studies on D-Ala-D-Lac Terminus Peptide Binding to Van-Group Antibiotics. The proposed research will: Demonstrate the versatility of CE in determining physicochemical parameters of antibiotics; Demonstrate high-throughput derivatization of receptors and ligands coupled to ACE; Provide for rapid synthesis and accurate analysis of multiple drug targets simultaneously; Require reduced sample volumes compared to complementary analytical techniques, and; Reduce sample waste and disposal.

描述（由申请人提供）：本研究的主要目标是开发毛细管电泳（CE）作为一种分析技术，用于研究抗生素万古霉素（Van）、瑞斯托菌素（Ris）和替考拉宁（Tel）的理化参数。其重点是展示原理并开发可用于分析受体-配体相互作用和微尺度反应的有用生物分析程序。抗生素耐药细菌的进化使得Van类抗生素的研究和开发成为研究重点的关键领域。因此，通过化学修饰来探讨这些抗生素的理化参数的研究是非常有必要的。这项工作将使用两种特定的 CE 技术：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/VIS) 检测方案以及高效液相色谱 (HPLC) 和酶动力学对小分子进行化学分离和鉴定。该研究将开发新的生物分析技术，并将重点放在检查与预防疾病有关的小生物分子，从而解决公共卫生问题。这种理解将加强​​ CE 的设计、准备和应用以解决许多健康相关问题的科学基础。 该研究的具体目标是 I. 利用柱上微反应器技术和 ACE 估计抗生素和配体之间的结合常数。二.使用柱上微反应器技术确定抗生素的电荷和 pl。三．使用反应性化学试剂检查抗生素衍生化的动力学。四．估计连接抗生素与肽的结合常数。 V.进行D-Ala-D-Lac末端肽与Van族抗生素结合的实验研究。 拟议的研究将： 展示毛细管电泳在确定抗生素理化参数方面的多功能性；展示与 ACE 偶联的受体和配体的高通量衍生化；同时提供多个药物靶标的快速合成和准确分析；与补充分析技术相比，需要减少样品量；减少样品浪费和处置。

项目成果

Applications of capillary electrophoresis to molecular recognition and analysis of in-capillary enzyme-mediated transformations.

毛细管电泳在毛细管内酶介导转化的分子识别和分析中的应用。

• DOI: 10.1201/9781420018066.ch3
• 发表时间: 2007
• 期刊: Advances in chromatography
• 作者: Zavaleta,Jose;Chinchilla,DinoraB;Brown,Abby;Ramirez,Alejandra;Calderon,Violet;Sogomonyan,Taguhi;Montes,Ruth;Gomez,FrankA
• 通讯作者: Gomez,FrankA

Multiple-injection affinity capillary electrophoresis to estimate binding constants of receptors to ligands.

多次注射亲和毛细管电泳来估计受体与配体的结合常数。

• DOI: 10.1007/s00216-005-0047-0
• 发表时间: 2005
• 期刊: Analytical and bioanalytical chemistry
• 影响因子: 4.3
• 作者: Chinchilla,Dinora;Zavaleta,Jose;Martinez,Karla;Gomez,FrankA
• 通讯作者: Gomez,FrankA

Multiple-injection affinity capillary electrophoresis to examine binding constants between glycopeptide antibiotics and peptides.

• DOI: 10.1016/j.chroma.2005.10.062
• 发表时间: 2006-02
• 期刊: Journal of chromatography. A
• 作者: J. Zavaleta;Dinora B Chinchilla;Karla Martinez;F. Gomez

Partial-filling affinity capillary electrophoresis techniques to probe the binding of glycopeptide antibiotics to D-Ala-D-Ala terminus peptides.

部分填充亲和毛细管电泳技术探测糖肽抗生素与 D-Ala-D-Ala 末端肽的结合。

• 发表时间: 2006
• 期刊: Journal of capillary electrophoresis and microchip technology
• 作者: Zavaleta,Jose;Chinchilla,DinoraB;Kaddis,CatherineF;Martinez,Karla;Brown,Abby;Gomez,Alvaro;Pao,Amaris;Ramirez,Alejandra;Nilapwar,Sanjay;Ladbury,JohnE;Gomez,FrankA
• 通讯作者: Gomez,FrankA

Estimation of binding constants between ristocetin and teicoplanin and peptides using on-column ligand derivatization coupled to affinity capillary electrophoresis.

使用柱上配体衍生化与亲和毛细管电泳结合来估计瑞斯托菌素和替考拉宁与肽之间的结合常数。

• DOI: 10.1007/s00216-004-2574-5
• 发表时间: 2004
• 期刊: Analytical and bioanalytical chemistry
• 影响因子: 4.3
• 作者: Azad,Maryam;Brown,Abby;Silva,Isba;Gomez,FrankA
• 通讯作者: Gomez,FrankA