Preparation of benzopyrene artifi cial antigen and mouse polyclonal antibody

With BaP as raw materials,the hapten was synthesised by nitration and reduction of the two step chemical reaction.The immune antigen(BSA-oABaP) and the detection antigen(OVA-oABaP) were prepared through conjugation of the hapten with BSA and OVA by using EDC method.By UV and SDSPAGE to identify coupling was successful.Then immuned the BALB/c mice,the polyclonal antiserum titer was detected by indirect ELISA,sensibility was identified by blocking ELISA.The results show that the BSA-oABaP wave moves to the right when compared with oABaP,and the moving speed of BSA faster than BSA-oABaP in the SDS-PAGE.That is to say coupling is successful.Indirect ELISA showed a high titer above 1:103of the antiserum of all the six BALB/c mouse.The sensibility of antiserum of No.6 mouse was the best,which IC 50 was 22.14 μg/mL and has good specificity.The experiments shows that we obtained the benzopyrene polyclonal antiserum with good sensibility and specificity.This polyclonal antibody may lay a foundation for the further benzopyrene ELISA rapid detection kit development.