Estimation of Percent Conjugation of Cypermethric Acid Hapten to BSA and Tetanus Toxoid Using 2,4,6 Trinitrobenzenesulfonic Acid Method

The use of immunoassays for analysis of small molecules has been extensive in endocrinology, clinical chemistry and other fields. However, small molecules such as pesticides are not able to induce immunogenic response by themselves but can generate immune response when conjugated to higher molecular weight compounds such as proteins. When the haptens are coupled to proteins the estimation of per cent conjugation is important as higher the concentration of hapten to protein more can be the specificity of developed antibodies. Here 2,4,6, trinitrobenzene sulphonic acid method was used to determine the per cent conjugation. Cypermethric acid, a metabolite of cypermethrin was used as hapten and was coupled to BSA and tetanus toxoid and the per cent conjugation was found to be 40% and 27 %, respectively.