Isolation and characterization of protein from frigate mackerel fish (Auxis thazard), green mussel (Perna viridis) and white shrimp (Penaeus merguensis) for production of isolate allergen

Abstract

Seafoods including fish, crustaceans and shellfish play an important role in human nutrition. However, it is also recognised as an important cause of food allergies, especially in coastal countries, including Indonesia. Treatments of these diseases still rely on avoiding the sources of the alergens, however it can also cause malnutrition which is also dangerous to health. To determine the specific food protein causing allergic reactions, cutie test was performed using isolate food protein. Until the present time, Indonesia still imports kits containing isolate proteins for food allergy diagnosis, therefore effort to locally produce the kit will reduce health cost for patients with allergic diseases. The objective of this study was to produce seafoods protein extract that is potential to be used as isolate of allergens for food allergy diagnosis. Two extract sarcoplasmic and myofibril protein were obtained from fish, shrimp and mussel by extraction in phosphate buffer with diference ionic strength Protein profile were then detected by means of sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE). IgE binding pattern was analyzed by using ELISA and immunoblotting using sera from 20 respondents with histories of food allergy. The result of the study showed that samples of seafood extract protein consist of several protein with various molecular weight in the range about 14 to 143 kDa. The component were identified as tropomyosin (34-38 kDa), parvalbumin (~12 kDa), myosin light chain (~17.5 kDa) and arginin kinase (40 kDa) which are major allergens in seafoods. Further analysis with ELISA showed that the seafood protein extracts contained several IgE-binding proteins. Immunoblot of three extract samples from seafoods showed that components identified as allergens to each allergic subject have different molecular weight. The results demonstrated that crude extract could be directly used as isolate allergen for diagnostic test allergy (skin prick test) without isolating the major allergen components.