Pooling of Monoclonal Antibodies for Rapid Detection of Commercially Important Fin Fish

• Other Authors: Zhu, Yaozhou (authoraut)
• Format: Others
• Language: English; English
• Published: Florida State University
• Subjects: Nutrition; Food; Exercise
• Online Access: http://purl.flvc.org/fsu/fd/FSU_migr_etd-8100

Due to the nutritional and health benefits, the consumption of fish as an important dietary protein source is increasing. However, the benefits are not always received by everyone. Fish is one of the eight major allergenic foods or food types and its prevalence in the United States was determined to be 0.4% of food-hypersensitive patients among the adult population. Approximately 1.1 million Americans suffer from various symptoms of fish allergy, and this number is increasing. Food manufacturers have been mainly depending on good manufacture practice (GMP) and hazards analyze and critical control point (HACCP) plan to avoid the presence of undeclared allergens in their products. Immunoassays using antibodies are widely accepted by regulatory agencies as a rapid and sensitive method for screening and monitoring substances in food and agricultural products. However, the vast species genetic variation of fish results in failure of identifying a single protein as a marker for fish protein detection. The specific objectives are: 1) to examine the selectivity of each MAbs 8F5, 2G9 and 2F3 by using indirect non-competitive ELISA(iELISA) and their antigenic proteins using western blot; 2) to develop a user friendly dot blot using pooled antibody for the detection of the presence of fish protein from all common food fish species; 3) to study the effects of storage and processing on the immunoreactivity thus the detestability of fish proteins using iELISA and dot blot. Three anti-fin fish MAbs (8F5, 2G9 and 2F3) were previously developed in our laboratory against cooked crude fish protein extract from red snapper, yellow fin tuna and swordfish, respectively. None of them was able to show the ideal pattern of recognition of the target fish species without cross reactivity or cross react with all fish species tested. But the cross reactivity pattern of each MAb complements to each other which may be utilized to recognize of a complete spectrum of most commercially important fish species. The overall goal of this research is to develop rapid user-friendly immunochemical detection methods using pooled MAb 8F5, MAb 2G9, and MAb 2F3 for the detection of fish proteins. The expected outcomes including: 1) Pooling of these three antibodies covered the immunoreactivity of commercially important fin fish and no cross reactivity with non-fish protein; the antigenic proteins of each MAb will also be identified; 2) pooling of MAbs would be effective for the detection of commercially important fish species using iELISA and dot blot assay; 3) iELISA and dot blot assay are effective and valid for fish detection under various storage and processing conditions. The pooled MAbs can be used as a standard reagent for detection of common fish species. Furthermore, a consumer-friendly immunoassay for the detection of the presence of fish could decrease the time of training experimental personnel. The research could provide a user-friendly tool not only to the regulatory agency for ensuring the enforcement of food laws but also the fish sensitive patients for risk reduction. === A Thesis submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Master of Science. === Spring Semester, 2013. === November 20, 2012. === Detection, Dot blot, ELISA, Fish, Pooled MAbs === Includes bibliographical references. === Yun-Hwa Peggy Hsieh, Professor Directing Thesis; Shridhar K. Sathe, Committee Member; Mary Ann Moore, Committee Member.