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Effects of Vinegar Treatment on Detectivity and Allergenicity of Finfish

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Effects of Vinegar Treatment on Detectivity and Allergenicity of Finfish Florida State University Libraries Electronic Theses, Treatises and Dissertations The Graduate School 2013 Effects of Vinegar Treatment on Detectability and Allergenicity of Finfish Ye Wang Follow this and additional works at the FSU Digital Library. For more information, please contact [email protected] THE FLORIDA STATE UNIVERSITY COLLEGE OF HUMAN SCIENCES EFFECTS OF VINEGAR TREATMENT ON DETECTABILITY AND ALLERGENICITY OF FINFISH By YE WANG 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 Degree Awarded: Summer Semester, 2013 Ye Wang defended this thesis on June 24th, 2013. The members of the supervisory committee were: Yun-Hwa Peggy Hsieh Professor Directing Thesis Shridhar Sathe Committee member Ming Cui Committee member The Graduate School has verified and approved the above-named committee members, and certifies that the thesis has been approved in accordance with university requirements. ii ACKNOWLEDGMENTS I would like to express my sincerest appreciation to my advisor Dr. Yun-Hwa Peggy Hsieh for all the opportunities she afforded throughout the past two years, without her support this work would not have been completed. Her constant inspiration and insightful comments were invaluable to my personal and professional development. I also would like to thank Dr. Shridhar Sathe and Dr. Ming Cui for being on my thesis committee despite their extremely busy schedule. Additionally, I would like to thank Dr. Jack Ofori, Dr. Yi- Tien Chen, Dr. Yuhong Wang, Behnam Keshavarz, and William Fredericks for their willingness to advise and help me in all areas throughout my course of study. And also thank you to my dear friends Yitong Zhao and Jingjie Xiao for their kindness and help in my life. Finally, I express my greatest thank to my mother Yonghua Wu for her unconditional love and support. Without her, I would not have had the opportunity to study in the U.S and have this wonderful life. iii TABLE OF CONTENTS List of Tables ................................................................................................................................. vi List of Figures ............................................................................................................................... vii Abstract .......................................................................................................................................... ix CHAPTER ONE: INTRODUCTION ..............................................................................................1 CHAPTER TWO: LITERATURE REVIEW ..................................................................................4 2.1 Fish Allergy ...................................................................................................................4 2.2 Fish Allergens ................................................................................................................5 2.2.1 Fish Parvalbumins ..............................................................................................5 2.2.2 Other Fish Allergens ..........................................................................................6 2.3 Effects of Processing on Food Proteins ..........................................................................7 2.3.1 Thermal Processing .............................................................................................7 2.3.2 Acid Processing .................................................................................................8 2.4 Detection Methods ..........................................................................................................9 2.4.1 Diagnosis of Food Allergies ...............................................................................9 2.4.2 Detection of Allergenic Foods ..........................................................................11 CHAPTER THREE: HYPOTHESES AND OBJECTIVES..........................................................13 3.1 Hypotheses ...................................................................................................................13 3.2 Objectives .....................................................................................................................13 CHAPTER FOUR: MATERIALS AND METHODS ...................................................................14 4.1 Materials ......................................................................................................................14 4.2 Methods ........................................................................................................................15 4.2.1 Sample Preparation ...........................................................................................15 4.2.2 Protein Extraction .............................................................................................16 4.2.3 Indirect ELISA ..................................................................................................16 4.2.4 Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Western Blot ..........................................................................................................17 CHAPTER FIVE: RESULTS AND DISCUSSION ......................................................................18 5.1 Effects of Different Types of Vinegar on the Detectability of Whiting .......................18 5.2 Effects of Vinegar Treatment Time on the Detectability of Three Common Species of Finfish ....................................................................................................................................19 5.3 Effects of Vinegar Treatment on Protein Banding Patterns of Finfish ........................20 5.4 Effects of Vinegar Treatment on Antigenic Protein of MAb 8F5 in Finfish ...............21 5.5 Effects of Vinegar Induced Chemical Reactions on the Detectability of Finfish ........22 5.6 Effects of Vinegar Treatment on the Allergenicity of Finfish .....................................24 5.7 Effects of Vinegar Induced Chemical Reactions on the Allergenicity of Finfish ........26 CHAPTER SIX: CONCLUSIONS ................................................................................................29 iv APPENDIX ...................................................................................................................................30 A. TABLES AND FIGURES ........................................................................................................30 REFERENCES ..............................................................................................................................48 BIOGRAPHICAL SKETCH .........................................................................................................56 v LIST OF TABLES Table 1. Fish allergens ...................................................................................................................30 Table 2. Total soluble protein concentration of whiting, cod and red grouper ..............................31 Table 3. Clinical features of patients .............................................................................................31 Table 4. Immunoreactivity of MAb 8F5 against vinegar-treated fish samples determined by iELISA ...........................................................................................................................................32 vi LIST OF FIGURES Figure 1. Effects of different types of vinegar on the detectability of whiting by MAb 8F5 based- iELISA. ..........................................................................................................................................32 Figure 2. Effects of vinegar treatment on the detectability of whiting, cod and red grouper by MAb 8F5-based iELISA ................................................................................................................33 Figure 3. SDS-PAGE profiles of vinegar-treated, water-treated and control samples of whiting, cod and red grouper .......................................................................................................................34 Figure 4. Antigenic protein banding patterns of vinegar-treated, water-treated and control samples of whiting, cod and red grouper .......................................................................................35 Figure 5. Study of the effects of vinegar induced chemical reactions on the detectability of whiting, cod and red grouper (Group A) .......................................................................................36 Figure 6. Study of the effects of vinegar induced chemical reactions on the detectability of whiting, cod and red grouper (Group B) .......................................................................................37 Figure 7. Study of the effects of vinegar induced chemical reactions on the detectability of whiting, cod and red grouper (Group C) .......................................................................................38 Figure 8. Screen for IgE-binding reactivity of raw and cooked salmon and cod ..........................39 Figure 9. Effects of vinegar treatment on the IgE-immunoreactivity of whiting by human plasma- based iELISA .................................................................................................................................40 Figure 10. Effects of vinegar treatment on the IgE-immunoreactivity of cod by human plasma- based iELISA .................................................................................................................................41
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