Studies On Extraction,Purification And Detecting Techniques Of Metapenaeusis Ensis Allergen

Shrimp and their products are more and more get the favor from people by their nutrition value and special flavour,however,they are the one of common foods that induce allergy,and threaten human health seriously,so the relevant research about shrimp allergen protein are carried out owing to their important economic and social benefits.Shrimp allergen protein by ammonium sulfate precipitation,isolated from salt-soluble protein of Metapenaeus ensis,and purified protein was gained by Sephadex G-100 gel column chromatography.The result showed: a single elution peak was gained by Sephadex G-100 gel column chromatography from ammonium sulfate precipitation with pH 6.54 and 40% saturation,and a single electrophoretic band was gained from this elution composition by SDS-PAGE,and this purified protein showed strong allergen activity by Dot-ELISA,and a novel protein with molecule weight 36.1 kDa was affirmed as the important allergen of Metapenaeus ensis.Balb/c mice(female,6-8 weeks)were immunized with purified Metapenaeu ensis allergen protein by short-term immunization,titre and cross reactive of antiserum were detected by indirect ELISA.The results showed that antiserum with the titre of 1 : 6400 was gained by short-term immunization with only 19 days,and this antiserum has no cross reactive against Southern American prawn,Oratosquilla kempi schmitt,Fenneropenaeus Chinensis,and polyclonal antibody of Metapenaeu ensis allergen was extracted from this antiserum by ammonium sulfate precipitation,DEAE-Cellulose DE-52 ion-exchange column chromatography and Sephadex G-150 gel column chromatography successively.A detecting technique of Metapenaeusis ensis allergen with the principle of indirect ELISA was designed in this content,and the detection interval is 0.32μg/mL-16 μg/mL.In this paper,Metapenaeu ensis allergen,polyclonal antaibody of mice anti Metapenaeu ensis allergen were gained and detecting technique of Metapenaeusisensis allergen was established,feasible means was proposed on the application of detecting allergen of shrimp and their products.