Purification And Immunological Identification Of Major Allergens Of Eriocheir Sinensis

Type of food allergy as a food-borne disease has aroused widespread concern. As one of three categories for 21th century prevention and treatment disease, allergic diseases has been included in the focus by World Health Organization, and it become a major worldwide problem of hygiene. Type of food allergy, a common allergic disease could cause the main clinical symptoms such as dizziness, headache, chest tightness, nausea, vomiting, skin itching, shock and even death. Therefore, in order to protect people's lives and health, improve the people's quality of life, carrying out researches on the food (especially seafood) allergens posses huge economic benefits and social benefits.Crustaceans such as shrimps and crabs and their products, one of high quality animal proteins sources, is also one of eight categories which easily leading to food allergy.With the development of international trade,crustaceans were widely applied in food. However, edible crab of the potential harm causing allergies is more deserving of attention.This paper was based on the common crab--Eriocheir sinensis in southern of China. This paper mainly focused on crab allergen separation,purification and immunodetection, the results are as follows:First, SDS-PAGE was applied to separate total protein from the crab extract,SDS-PAGE spectrum showed that the proteins of Eriocheir sinensis were comprised of at least twenty discrete protein bands with the molecular weight ranging from15.7 KD-118 KD.The Eriocheir sinensis allergens were characterized by Western blotting using 7 seras from patients who were allergic to the crab. There are 9 protein bands that can bind with patients'IgE,detected from crab extract by Western blotting.Their molecular weights were: 29 KD, 34 KD, 35 KD, 41 KD, 48 KD, 53 KD, 60 KD, 66 KD and 76 KD. Among them, 4 protein bands of 35 KD, 41 KD, 48 KD and 66 KD have high positive rate, that may be the major allergens of Eriocheir sinensis.Second,the crab allergen proteins were gradually purificatied by salting out with the ammonium sulphate,DEAE-52 ion-exchange chromatography and Sephadex G-75 gel column chromatography.The pure allergen protein which has a high reactivity was analysised and identificatied by Western-blotting and ELISA.In this study,several major allergens were purificatied and identified from Eriocheir sinensis.It will be used as a base for further study on structure and function of crab allergens, As a specific antigen tested by ELISA,it has a good reaction to the specific IgE from patients who are allergic to the crab.It will also be used as a base for the specific diagnosis and immunotherapy in crab hypersensitivity disease and further development of immune reagent desensitization.