Abstract: 【Aim】 To obtain the recombinant allergen CPH30 in pupae of the silkworm, Bombyx mori, and to detect its immunogenicity. 【Methods】 The potential allergenicity of CPH30 protein was analyzed with proteomic methods. The CPH30 gene was synthesized by chemical method and introduced into pET-28a vector. The expression of the recombinant CPH30 was induced by IPTG. The allergenic activity of CPH30 protein was detected by Western blot. The potential B cell epitopes were analyzed using DNAStar. 【Results】 After induction with IPTG, CPH30 protein was largely expressed in Escherichia coli Rosetta and purified by affinity chromatography. The molecular weight of the recombinant CPH30 protein is 25 kD. The CPH30 protein show a high IgE binding activity with the serum from silkworm-allergic patients, indicating that the recombinant CPH30 protein has immunogenicity. Prediction result using DNAStar software showed that the potential B cell epitopes of CPH30 are located between the 57th-69th and 150th-158th amino acids .【Conclusion】 The purified CPH30 has high purity and immunogenicity. This study lays a foundation for the specific diagnosis, vaccine treatment and further experimental studies of allergic diseases caused by silkworm pupae.

Key words: Bombyx mori, CPH30, allergen, immunological identification, B cell epitope, immunogenicity