Purification And Identification Of Major Allergens Of Metapenaeus Ensis

Food allergy, a special type of foodborn disease, is currently of concern in the world. Allergic disease is regard as one of three dieases that need prevention and cure in 21st century. Food hypersensitivity is a normal allergic dieases, which can provokes dizziness, headache, chest distress, itch of skin and so on. So it is necessary to study the food allergens, especially seafood allergens. It has tremendoes economic and social benefits to protect health of people, and improve people's quality of life.Shrimps, crabs and other crustacean and their produce is one of the best animal proteins for human; it is also one of the eight kinds of foods that frequently causes allergy. As the development of world trade, shrimp was used more and more widely in all kinds of foods. The potential risk of shrimpallergy was worth to pay more attention. This paper was based on the common shrimp --- Metapenaeus ensis in southern china. This paper mainly focused on shrimp allergen separation purification and immunodetection, the results are as follows:First,SDS-PAGE was applied to separate total protein from the shrimp extract, SDS-PAGE spectrum showed the proteins of Metapenaeus ensis comprised of at least twenty discrete protein bands with the molecular weight ranging from12.1 KDa-105.6 KDa.The Metapenaeus ensis allergens were characterized by Western blotting using 12 seras from patients allergic to shrimp , There are 7 protein bands that can bind with patients, IgE were detected from shrimp extract by Western blotting,Their molecular weights were:36 KDa,46.2 KDa,60 KDa, 64.8 KDa,68 KDa,78 KDa and 88.5 KDa, Some patients also recognized the bands of 21.6KDa,Among them there are 4 protein bands of 36 KDa,46.2 KDa,60 KDa and 68 KDa have high positive rate, they may be the major allergens of Metapenaeus ensis.This results were different with the others in our country and abroad,It may be related to the different types of shrimp, their living environment is inconsistent in every region and the allergic patients, individual differences.Second, the shrimp allergen proteins were gradually purificatied by salting out with the ammonium sulphate, DEAE-52 ion-exchange chromatography and Sephadex G-100 gel column chromatography. The pure allergen protein was analysised and identificatied by western-blotting, dot-ELISA and ELISA, which has a high reactivity.In this study, several major allergens were purificatied and identified from Metapenaeus ensis. It will be used as a base for further study on structure and function of shrimp allergens, In ELISA it as a specific antigen has a good reactive to the specific IgE from patients who allergic to shrimp. It will also be used as a base for the specific diagnosis and immunotherapy in shrimp hypersensitivity disease and further development of immune reagent desensitization...