Capillary Electrophoresis: Bioanalytical Applications

描述（由申请方提供）：本研究的广泛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（货车）、利托那韦（Ris）和替考拉宁（Tel）的理化参数。它的重点是展示原理和开发有用的生物分析程序，可应用于受体-配体相互作用和微尺度反应的分析。抗生素耐药菌的进化使得Van类抗生素的研究和开发成为研究热点。因此，通过化学修饰探索这些抗生素的理化参数的研究是非常必要的。两个具体的CE技术将在这项工作中使用：柱上微反应器技术和亲和毛细管电泳（ACE）。在这项研究中使用的技术包括化学修饰和表征，化学分离和鉴定的小分子CE利用激光诱导荧光（LIF）和紫外/可见光（UV/维斯）检测方案和高效液相色谱（HPLC），和酶动力学。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的设计，准备和应用对一系列健康相关问题的科学基础。 本研究的具体目的是：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.用柱上微反应器技术测定抗生素的电荷和pI。三.使用反应性化学试剂检查抗生素衍生化的动力学。四.估计连接的抗生素与肽的结合常数。进行D-Ala-D-Lac末端肽与VAN类抗生素结合的实验研究。 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，需要减少的样品体积;减少样品浪费和处置。

DESCRIPTION (provided by applicant): The broad goals of this research are to 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.