A new detection technique for fluoroquinolone-conjugated proteins by high performance liquid chromatography with UV/fluorescence detectors

Abstract

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 toot for the detection drug-carrier-conjugated antigens, especially for FQs-BSA-conjugates during the production of anti-FQs monoclonal antibodies.