Submitted to: Analytica Chimica Acta
Publication Type: Peer reviewed journal
Publication Acceptance Date: 10/9/2000
Publication Date: 10/12/2001
Citation: Interpretive Summary: In these studies, computers were used to produce models (pictures) of chemicals used in the production of antibodies to nicarbazin, a drug (coccidiostat) used to control diseases in poultry. Antibodies are molecules that are found in the blood that have the ability to specifically bind other molecules such as drugs, bacteria or viruses. The models we developed assisted us in defining a chemical (hapten) that was used to produce antibodies to nicarbazin. In this article we describe how we made the hapten. The results described here can be used to help make an antibody-screening test for the detection of nicarbazin in poultry feeds. This will allow the level of nicarbazin in poultry feeds to be monitored assuring the production of healthy poultry and proper use of nicarbazin.
Technical Abstract: Based on insights obtained from computer assisted molecular modeling, p- nitrosuccinanilic acid was selected as a viable candidate to use as a hapten to produce antibodies to 4,4'-dinitrocarbanilide (DNC), the active component of the coccidiostat nicarbazin. These antibodies would be used to quantify nicarbazin in feeds and tissues. The synthesis of p-nitrosuccinanilic acid is presented here. Due to the lack of water solubility of nicarbazin, a discussion is presented concerning the solubility of nicarbazin as it pertains to the immunoassay. DMF and ACN were added to the assay buffer to make standard solutions of nicarbazin, and facilitate the use of the competition enzyme-linked immunosorbent assays (ELISAs). Monoclonal antibodies (Mabs) were isolated that compete with nicarbazin, and they were isotyped as IgM.