Preparation and Purfication of High Specific IgG Antibody for the Affinity Chromatograpy of Chloramphenicol

Objective To prepare a self-made immunoaffinity chromatography column for the high purified anti-Chloramphenicol polyclonal antibody and investigate its biological activity.Method Purification of antibodies with ammonium sulphate fractional precipitations was conducted initially.Bis(3-aminopropg) amine was used as a spacer between gel and Chloramphenicol(CAP) ligand of affinity chromatography column to purify and differentiate the specific IgG for anti-CAP antibody from other crude IgG.The activity of antibody purified by affinity chromatography was determined by ELISA.Results The target proteins were precipitated by saturated ammonium sulfate,and most other proteins can be removed.Highly purified rabbit anti-chloramphenicol antibody can be obtained by the self-prepared immunoaffinity chromatography column.The biological activity of the target protein was 4 times higher than that before purification.Cross reaction showed that the IgG was highly specific to CAP,and litter response to other selected antimicrobial agents.Conclusion The established method can be used to obtain purified specific IgG from Chloramphenicol antibody,which may lay a foundation for the immunological examination of Chloramphenicol.