Preparation of Ciprofloxacin Derivative and its Antibody Specificity
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Abstract:
The preparations of the ciprofloxacin (CPLX) carboxyl derivative and its antibody were studied in this paper, and the antibody specificity of this derivative was determined. CPLX derivatives (CPLX-COOH) were synthesized with CPLX and 3-bromopropionic acid, and purified by preparation of liquid phase. Infrared spectroscopy (IR), electrospray ionization mass spectrometry (ESI-MS), and nuclear magnetic resonance (NMR) spectroscopy were used to identify the CPLX derivative. CPLX-COOH was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to prepare the immune antigen CPLX-COOH-BSA and coating antigen CPLX-COOH-OVA, which were identified using ultraviolet spectroscopy. The BALB/c mice were immunized with CPLX-COOH-BSA to obtain a polyclonal antibody directed against the CPLX derivative with high titer. The optimum concentration of the coating antigen and the optimum dilution of the antibody were determined using the chessboard method, and the indirect competitive enzyme-linked immunosorbent assay (ELISA) was established. The half-maximal inhibitory concentration (IC50) and the limit of detection (LOD) of CPLX-COOH were 228.56 ng/mL and 25.527 ng/mL, respectively. This antibody had a broad specificity and cross reactivity with multiple fluoroquinolones, and could be used to detect several fluoroquinolone residues.
