ISSN print edition: 0366-6352 ISSN electronic edition: 1336-9075 Registr. No.: MK SR 9/7 Published monthly

Preparation and characterization of ultrasensitive and specific polyclonal antiserum against ciprofloxacin based on cationized bovine serum albumin

Sheng-Liang Deng, Ping Li, Hong-Bin Liu, and Shu-Ming Yang

Institute of Quality Standard and Testing Technology for Agricultural Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

E-mail: yangshumingcaas@sina.com

Abstract: An ultrasensitive polyclonal antiserum against ciprofloxacin (CPFX) was developed by introducing cationized bovine serum albumin (BSA) as the carrier. Conjugates used as immunogens were synthesized using CPFX coupled to BSA cationized with ethylenediamine and hexamethylenediamine, namely CPFX-eBSA and CPFX-hBSA. The results showed that the antisera immunized with CPFX-hBSA exhibit an improved sensitivity and specificity compared with those immunized with CPFX-eBSA. Under the conditions of optimal heterologous enzyme-linked immunosorbent assay, the optimal antiserum yielded an IC₅₀ of 0.097 ng mL⁻¹, which proves it to be more sensitive than any CPFX antibody. Cross-reactivities with norfloxacin, enrofloxacin, and ofloxacin were < 3.0 %. No cross-reactivities with penicillin and gentamicin were found. The limit of detection was 0.01 ng mL⁻¹, which is far below the maximum residue level established by the EU regulation, suggesting the great potential of the presented antiserum for quantitative assays of CPFX in samples.

Keywords: ciprofloxacin – cationized BSA – polyclonal antiserum – heterologous ELISA

Full paper is available at www.springerlink.com.

DOI: 10.2478/s11696-014-0593-z

Chemical Papers 68 (11) 1505–1513 (2014)