Characterization of Anti-Chloramphenicol Antibodies by Enzyme-Linked Immunosorbent Assay

Abstract

Polyclonal antibodies against conjugates of chloramphenicol succinate and chloramphenicol base with proteins were obtained and characterized in direct ELISA. Antiserum against a conjugate of chloramphenicol (CAP) base with BSA (direct coupling) was very specific and showed cross-reactivity only with CAP succinate (11.3%) and CAP base (4.6%); whereas, antisera against a conjugate of CAP succinate with a protein recognized CAP succinate strongly as an initial compound. In direct ELISA, antisera against a conjugate of CAP succinate with KLH (homologous assay) and CAP base with BSA (heterologous assay) showed similar sensitivity: IC₅₀ were 1.3 and 1.5 ng mL⁻¹, respectively. Applicability of the immunoreagents obtained was shown in the analysis of CAP residues in milk (3.5% fat content). Detection limit of 0.3 ng mL⁻¹ was obtained for milk diluted 5 times.

Keywords:

• Antibodies
• Chloramphenicol
• Enzyme-linked immunosorbent assay

Notes

*Cross reactivity values for other antibiotics (ampicillin, gentamicyn, penicillin, streptomicyn, erithromicyn) were less then 0.1%.