Bioanalytical Applications of Capillary Electrophoresis

描述（由申请人提供）：本研究的主要目标是进一步发展毛细管电泳（CE）作为一种分析技术来研究糖肽类抗生素万古霉素（Van）、瑞斯托司汀（Ris）和替科planin （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，可以应用于分析受体-配体相互作用和微尺度反应。抗菌剂耐药性的发展是一个世界性的问题，它继续危及用于治疗许多传染病的新药的临床效果。因此，有必要通过化学修饰对这些抗生素的理化性质进行研究。在拟议的研究中，两种特定的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，化学分离和小分子的CE鉴定，利用激光诱导荧光（LIF）和紫外/可见（UV/VIS）检测方案和高效液相色谱（HPLC）。这项研究将开发新的生物分析技术，并将重点研究与疾病预防有关的小生物分子，从而涉及公共卫生问题。这种理解将加强CE的设计、制备和应用于一系列健康相关问题的科学基础。该研究的具体目的是：1 .将列联抗生素衍生化反应与ACE结合。2。检查与ACE耦合的柱上配体衍生化反应。3。利用多次注射ACE （MIACE）分析配体与糖肽类抗生素的结合。四、优化流动-部分填充ACE （FTPFACE）分析配体与抗生素结合的条件。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；展示高通量衍生化受体和配体偶联到ACE；提供药物靶点的快速合成和准确分析；与其他分析技术相比，需要更少的样本量；减少样品的浪费和处理。

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.