Bioanalytical Applications of Capillary Electrophoresis

描述(申请人提供)：这项研究的主要目标是进一步发展毛细管电泳法(CE)作为一种分析技术，研究糖肽抗生素万古霉素(Van)、瑞沙西汀(Ris)和替考拉宁(Tei)及其衍生物的物理化学性质。它的重点是演示原理和开发有用的分析技术，这些技术可以应用于受体-配体相互作用和微型反应的分析。抗菌药耐药性的产生是一个世界性的问题，它继续损害用于治疗许多传染病的新药的临床有效性。因此，需要研究通过化学修饰来探索这些抗生素的物理化学性质。在拟议的研究中，两种特定的毛细管电泳法将是：柱上微反应器技术和亲和毛细管电泳法。研究中使用的技术包括化学修饰和表征、利用激光诱导荧光(LIF)和紫外可见(UV/VIS)检测方案以及高效液相(HPLC)的CE化学分离和鉴定小分子。这项研究将开发新的生物分析技术，并将重点放在检查涉及疾病预防和公共卫生问题的小生物分子上。这一理解将加强CE的设计、准备和应用的科学基础，以解决一系列与健康有关的问题。本研究的具体目的是：1.将抗生素衍生化反应与ACE偶联。II.考察与ACE偶联的柱上配体衍生化反应。利用多次注射血管紧张素转换酶(MIACE)分析配体与糖肽类抗生素的结合。优化流通式部分填充ACE(FTPFACE)条件，分析配体与抗生素的结合情况。拟议的研究将：展示CE在确定抗生素的物理化学参数方面的多功能性；展示与ACE连接的受体和配体的高通量衍生化；提供药物靶标的快速合成和准确分析；要求与其他分析技术相比减少样品体积；减少样品浪费和处置。

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.