Preparation of monoclonal antibody against Bt Cry1Ac by using synthetic peptide as Hapten.

In order to prepare monoclonal antibody Bt Cry1Ac,we got the amino acid sequence of Bt Cry1Ac protein from NCBI,and synthesized specific Bt Cry1Ac peptide,according to its antigenicity,hydro-philicity and accessibility on the basis of its structural features by computer assisted analysis.The peptide was linked with carrier keyhole limpet hemocyanin(KHL),and then used for immunized mouse as antigen.34 strains of hybridoma,which can produce monoclonal antibodies to the peptide,have been obtained by cell fusion technique.One hybridoma strain(6G9),which was specific to natural Bt Cry1Ac was selected by indirect ELISA and Western blot.The result showed that the monoclonal antibody of hybridoma strain(6G9) can react specifically with the synthetic peptide and Bt Cry1Ac protein,but can not react with Bt Cry1Aa and Bt Cry1Ab proteins by ELISA and Western blot.This result of experiment showed that the monoclonal antibody of hybridoma strain(6G9)can distinguish transgenic Bt CrylAc cotton and general cotton by ELISA,and it can recognize specifically Bt Cry1Ac protein in cotton.