Immunodetection of Allergens from Mullet (Mugil Cephalus) and Salmon (Salmo Salar)

• Other Authors: Keshavarz, Behnam (authoraut), Hsieh, Peggy (professor co-directing dissertation), Rao, Qinchun, 1974- (professor co-directing dissertation), Chase, P. Bryant (university representative), Arjmandi, Bahram H. (committee member), Kim, Jeong-Su (committee member), Sathe, Shridhar K. (committee member), Florida State University (degree granting institution), College of Human Sciences (degree granting college), Department of Nutrition, Food and Exercise Sciences (degree granting departmentdgg)
• Format: Doctoral or Postdoctoral Thesis
• Language: English
• Published: Tallahassee, Florida: Florida State University 2017
• Subjects: Food; Medical sciences; Immunology; Salmo salar
• Online Access: https://diginole.lib.fsu.edu/islandora/object/fsu%3A507690/datastream/TN/view/Immunodetection%20of%20Allergens%20from%20Mullet%20%28Mugil%20Cephalus%29%20and%20Salmon%20%28Salmo%20Salar%29.jpg

The presence of misbranding and undeclared allergenic residues was the number one cause of food recalls in the United States, between 2012 to 2016. In this study, parvalbumin was used as a model to study the matrix effect on thermostability of this protein. In general, parvalbumin is thermostable; however, its thermostability varies among different fish species. Identification of new fish allergens is also another important factor for the development of immunoassays to determine allergen-specific IgE antibodies. The specific objectives of this study were to: 1) study the matrix effect on thermostability of parvalbumin from mullet and salmon using three sample models, and 2) identify new potential fish allergen(s) and evaluate the in vitro pepsin digestion stability of suspect allergen(s). To fulfill objective 1, three sample systems were studied: soluble protein extracts from hot smoked samples, protein extracts (PE), and purified parvalbumin (PP). The PE and PP samples were heated for 0, 2, 5 and 8 min at 100 °C, respectively. BCA assay, SDS-PAGE, indirect non-competitive ELISA (inELISA), and Western blot (WB) were used to study the relative protein solubility (RPS), molecular integrity, relative immunoreactivity (RI), and antigenicity of parvalbumin in PE and PP samples., respectively. The amino acid (AA) sequence of mullet parvalbumin was determined using LC-MS/MS. Overall, RPS of PE samples heated for 8 min, compared with unheated samples, were significantly decreased (P < 0.05) in both species; however, no significant decreases (P < 0.05) were observed in RPS of PP samples. From SDS-PAGE, parvalbumin color intensity in PP model did not change over the heating time. Whereas, it was decreased in the salmon smoked and heated PE. From ELISA, in mullet PE, RI of heated samples did not significantly change (P > 0.05). However, RI of salmon PE significantly decreased (P < 0.05) as a function of heating time. The RI of mullet PP was not significantly different over the heating time (P > 0.05), while ...