Partial Purification And Identification Of Major Allergens In Alternaria Alternata

Background Alternaria alternata is one of the most important and common fungal allergens in the world. Many articles about purification and identification of Alternaria allergens have been reported abroad so far. But few have been reported in our country.Objective Our research is to extract allergens of Alternaria alternata, to identify and partially purify the primary and the secondary allergens of Alternaria alternata, to provide theoretical basis for the standardization of fungal allergens.Methods On the basis of our previous investigation on aero-allergens in Nanchan city, Jiangxi province, we selected Alternaria alternata as our research subject. The Alternaria alternate allergens were extracted by different extraction methods , the protein allergen components were isolated by means of SDS-polyacrylamide gel electrophoresis (SDS-PAGE) . And its primary and secondary allergen proteins have been identified by Western-blotting. Then, primary and secondary allergen proteins of Alternaria alternata were purified by DEAE Cellulose-52 Ion Exchange Chromato-graphy (IEC).Their bioactivities were detected by Western-blotting.Results Several protein bands from Alternaria alternata extract have been isolated by means of SDS-polyacrylamide gel electro-phoresis (SDS-PAGE).The results show that the antigens with themolecular weight of 40KD and 45KD are the primary allergens of Alternaria alternata;Unique allergen protein bands of the Alternaria alternata were identified , whose molecular weights are 40KD and 45KD. And its primary and secondary allergen proteins have been identified by Western blotting.Then the primary and secondary allergen proteins of Alternaria alternata were purified by DEAE Cellulose-52 Ion Exchange Chromatography (IEC). Its bioactivity was detected by Western-blotting. Its molecular weight determined by SDS-PAGE was 40K Dalton and 45K Dalton.Conclusion The major allergens represent the main allergic components of Alternaria alternata. The isolation, identification and purification of the primary antigens of Alternaria alternata will provide theoretical basis for the development of standardized fungal allergen and explore safer and more effective diagnosis and treatment methods for the fungal allergic diseases.