Preparation and purification of monoclonal antibodies against chloramphenicol

Monoclonal antibodies (McAbs) against chloramphenicol (CAP) were produced to detect CAP residues, which could be toxic and possesses a potential threat to human health. The CAP-BSA conjugate was obtained by bovine serum albumin (BSA) coupled with CAP, and used to immunize the mice. The splenocytes from the immunized mice were fused with mouse myeloma cells SP2/0 to form hybridoma, which may secrete McAbs against CAP. Hybridomas 1D₁ and 3G₁₂ secreting McAbs against CAP were obtained by screening. Ascites containing McAbs were prepared by injecting 1 x 10⁶ cells of hybridoma 1D₁ and 3G₁₂ into the abdomen of mice. Protein A affinity chromatography was used to purify McAbs against CAP in a single chromatographic step with recovery yield above 80% and purity above 95% and full recovery of antibody activity. Experiments showed that McAb 3G₁₂ was highly specific for CAP and had no cross-reactivity with analogues which have a structure similar to CAP. The IC₅₀ value was 50.8 ng/mL.