[Conjugation and immunogenic evaluation of complete immunogen for the small molecular environmental pollutants 2,4-D]

Conjugation of complete immunogen for 2,4-dichlorophenoxyacetic acid (2,4-D) was studied. 2,4-D was cross-linked to bovine serum albumin (BSA, carrier) by 1-ethyl-3-(3-dimethyl- aminopropyl) carbodiimide hydrochloride (EDC). The conjugation reaction was found to be more effective at 4 degrees C and incubated for 18 hour. 2,4-D was dissolved in 0.05 mol/L phosphate buffer between 10.0 - 12.0 mg/mL concentration, pH was adjusted to be 5.4 - 6.1. When the weight of added EDC was below 12 mg, the more EDC was added the higher substitution degree complete immunogen was synthesized. Complete immunogen of various substitution degree (2,4-D: protein) were applied to immunize balb/c mice. The conjugates of 2,4-D and poly-L-lysine was applied as coating antigen. It was experimentally found that complete immunogen of substitution degree 12 and 18 are more immunogenic than that of substitution degree 6 and 25. None-specific adsorption between antiserum that was produced by complete immunogen of substitution degree 18 and coating antigen was very weak, and the antiserum contained more 2,4-D specific antibody. It could be used as the immunogen for the preparation monoclonal antibody.