利用高效液相層析儀搭配紫外光和螢光檢測器檢測氟化奎林酮類藥物螯合蛋白之新檢測技術   全文下載

New Detection Technique for Fluoroquinolone-Conjugated Proteins by High Performance Liquid Chromatography with UV/Fluorescence Detectors

本研究擬開發一套簡單、快速及可信賴的檢測氟化奎林酮類藥物（f1uoroquinolones，FQs）以N-hydroxysuccinimide ester方法結合牛血清白蛋白（bovine scrum albumin，BSA）之技術。此技術係依據FQs和BSA的不同吸收特性，其中FQs於高效液相層析儀搭配紫外光和螢光檢測器可被檢測出，但BSA卻無法被螢光檢測器所檢測出。結果顯示，利用此方法可以有效地檢測出FQs-BSA的藥物螯合蛋白，而此方法不僅可以用來檢測FQs和BSA，也可用來評估FQs-BSA的螯合程度。因此，此檢測技術為一種藥物-載體結合抗原有效的檢測工具，尤其在抗FQs類單株抗體製作過程中，來評估FQs-BSA的結合物。

This study was aimed to develop a simple, fast, and reliable technique to detect fluoroquinolones (FQs) -conjugated bovine serum albumin (BSA). Four tested FQs, enrofloxacin, ofloxacin, danofloxacin, and orbifloxacin, were conjugated with BSA by following the N-hydroxysuccinimide ester method. The technique was designed according to the different absorption characteristics of the FQs and BSA; FQs can be detected by both UV and fluorescence detectors, but BSA can only be detected by UV. The results demonstrated that the developed method was efficient in detecting FQs-BSA conjugates. In addition, the method not only trace the FQs and BSA conjugation responses but also can be used to estimate the level of FQs-BSA conjugation. Therefore, this technique is a valuable tool for the detection drug-carrier-conjugated antigens, especially for FQs-BSA-conjugates during the production of anti-FQs monoclonal antibodies.