A new method for the purification of shrimp allergenic protein Pen a 1 was developed.The epitope peptide（aa 247 to 284） of the Pen a 1 was synthesized by standard Fmoc and conjugated to KLH and BSA to get the artificial immune antigen（peptide-KLH） and the coating antigen（peptide-BSA），respectively.New Zealand white rabbits were immunized with immunogen peptide-KLH to produce antisera and the antisera were purified by bitterness-ammonium sulfat and HiTrap rProtein A FF.The pure antiserum was coupled to CNBr-Activated Sepharose 4B.The titer of purified antibody was 2.05×106 by indirect ELISA and the IC50 was 0.21 mg/L by indirect competitive ELISA method.There was no cross-reactivity between the antibody and the proteins from shrimp except for Pen a 1 by indirect competitive ELISA method.The coupling efficiency of the antibody with CNBr-Activated Sepharose 4B was 90.76% and the column capacity for 1 mL immunosorbents was 2.84 mg Pen a 1.The recoveries of the immunoaffinity column were in the range of 89.6%-93.6%.The service life of the immunoaffinity column was 4 cycles.The SDS-PAGE and Western Blot results displayed that the purified Pen a 1 had high immunogenicity.