Digital Commons Network^™

Safety Assessment Of Novel Foods And Food Proteins, Niloofar Moghadam Maragheh

Department of Food Science and Technology: Dissertations, Theses, and Student Research

The global food market needs to grow and supply food demand to feed the growing world population. Alternative food proteins, including novel sources of safe foods and ingredients, are the candidates that could provide more environmentally sustainable choices, animal welfare, and consumers health. Novel foods and food proteins must undergo premarket safety evaluations including allergenicity assessment to reduce the risk of cross-reactivity with known allergens and uncharacterized risk to food allergic individuals. This research addressed the safety assessment of some novel foods and food ingredients using the study of stability of proteins in pepsin and sequence identity analysis in the …

Novel Sources Of Food Allergens, Lee Palmer

Department of Food Science and Technology: Dissertations, Theses, and Student Research

Technological advancement and globalization have led to the spread of foods to countries where the food does not yet have a documented history of consumption, in other words, novel foods. Novel foods also encompass truly novel foods, foods that have been processed in a novel manner, and novel means of exposure. With novel foods comes the potential of food allergens that pose an uncharacterized risk to those with food allergies. Food allergies are an increasingly important facet of public health. Therefore, a deeper understanding of novel foods as well as methods to evaluate consumers’ potential risk is necessary. Literature reviews …

Development Of A Sandwich Elisa Targeting Cashew Ana O 2 And Ana O 3, Morganne Schmidt

Department of Food Science and Technology: Dissertations, Theses, and Student Research

Cashew nut is the second leading tree nut allergy in the US. Cross-contact of cashew nut poses potential food safety risks for individuals with cashew allergies. Highly-processed foods, such as HTST/UHT cashew milks may lead to problems in cashew protein detection by current allergen detection methods. Therefore, the aim of this study was to develop a robust sandwich ELISA for detection of highly processed cashew proteins. Commercial cashew ELISAs were evaluated for their robustness and sensitivity in detecting cashew milk cashew protein. After unreliable results were determined, cashew Ana o 2 (11S) and Ana o 3 (2S) were semi-purified using …

Purification And Characterization Of Naturally Occurring Post-Translationally Cleaved Ara H 6, An Allergen That Contributes Substantially To The Allergenic Potency Of Peanut, Govardus A. H. De Jong, Shyamali Jayasena, Phil Johnson, Justin Marsh, Danijela Apostolovic, Marianne Van Hage, Julie Nordlee, Joe Baumert, Steve L. Taylor, Camille Roucairol, Harmen De Jongh, Stef J. Koppelman

Department of Food Science and Technology: Faculty Publications

The 2S albumin Ara h 6 is one of the most important peanut allergens. A post-translationally cleaved Ara h 6 (pAra h 6) was purified from Virginia type peanuts, and the cleavage site was mapped using high-resolution mass spectrometry. Compared to intact Ara h 6, pAra h 6 lacks a 5-amino acid stretch, resembling amino acids 43−47 (UniProt accession number Q647G9) in the nonstructured loop. Consequently, pAra h 6 consists of two chains: an N-terminal chain of approximately 5 kDa and a C-terminal chain of approximately 9 kDa, held together by disulfide bonds. Intermediate post-translationally cleaved products, in which this …

Release Of Major Peanut Allergens From Their Matrix Under Various Ph And Simulated Saliva Conditions—Ara H2 And Ara H6 Are Readily Bio-Accessible, Stef J. Koppelman, Mieke Smits, Monic Tomassen, Govardus A.H. De Jong, Joseph L. Baumert, Steve L. Taylor, Renger Witkamp, Robert Jan Veldman, Raymond Pieters, Harry J. Wichers

Department of Food Science and Technology: Faculty Publications

The oral mucosa is the first immune tissue that encounters allergens upon ingestion of food. We hypothesized that the bio-accessibility of allergens at this stage may be a key determinant for sensitization. Light roasted peanut flour was suspended at various pH in buffers mimicking saliva. Protein concentrations and allergens profiles were determined in the supernatants. Peanut protein solubility was poor in the pH range between 3 and 6, while at a low pH (1.5) and at moderately high pHs (>8), it increased. In the pH range of saliva, between 6.5 and 8.5, the allergens Ara h2 and Ara h6 …

Endurance Exercise Increases Intestinal Uptake Of The Peanut Allergen Ara H 6 After Peanut Consumption In Humans, Lonneke M. Janssenduijghuijsen, Klaske Van Norren, Sander Grefte, Stef J. Koppelman, Kaatje Lenaerts, Jaap Keijer, Renger F. Witkamp, Harry J. Wichers

Department of Food Science and Technology: Faculty Publications

Controlled studies on the effect of exercise on intestinal uptake of protein are scarce and underlying mechanisms largely unclear. We studied the uptake of the major allergen Ara h 6 following peanut consumption in an exercise model and compared this with changes in markers of intestinal permeability and integrity. Ten overnight-fasted healthy non-allergic men (n = 4) and women (n = 6) (23 +/- 4 years) ingested 100 g of peanuts together with a lactulose/rhamnose (L/R) solution, followed by rest or by 60 min cycling at 70% of their maximal workload. Significantly higher, though variable, levels of Ara h 6 …

Allergenicity Attributes Of Different Peanut Market Types, Stef J. Koppelman, Shyamali Jayasena, Dion Luykx, Erik Schepens, Danijela Apostolovic, Govardus A.H. De Jong, Thomas G. Isleib, Julie A. Nordlee, Joseph L. Baumert, Steve L. Taylor, Hsiaopo Cheng, Sohelia Maleki

Department of Food Science and Technology: Faculty Publications

Four different market classes of peanut (Runner, Virginia Spanish, and Valencia) are commonly consumed in Western countries, but for some consumers peanuts are a main cause of food-induced anaphylaxis. Limited information is available on the comparative allergenicity of these distinct market classes. The aim of this study was to compare allergenicity attributes of different peanut cultivars. The protein content and protein profiles were highly comparable for all tested cultivars. All cultivar samples contained the major allergens Ara h 1, Ara h 2, Ara h 3 and Ara h 6, as assessed by SDS-PAGE and RP-HPLC, although some minor differences in …

Ara H 6 Complements Ara H 2 As An Important Marker For Ige Reactivity To Peanut, Audrey E. Koid, Martin D. Chapman, Robert G. Hamilton, Ronald Van Ree, Serge A. Versteeg, Stephen C. Dreskin, Stef J. Koppelman, Sabina Wünschmann

Department of Food Science and Technology: Faculty Publications

The similarities of two major peanut allergens, Ara h 2 and Ara h 6, in molecular size, amino acid sequence, and structure have made it difficult to obtain natural Ara h 6 free of Ara h 2. The objectives of this study were to purify natural Ara h 6 that is essentially free of Ara h 2 and to compare its IgE reactivity and potency in histamine release assays to Ara h 2. SDS-PAGE of the highly purified allergen (<0.01% Ara h 2) revealed a single 14.5kD band and the identity of Ara h 6 was confirmed by LC-MS/MS. Ara h 6 showed a higher seroprevalence in chimeric-IgE ELISA (n=54), but a weaker biological activity in basophil histamine release assays than Ara h 2. Purified Ara h 6 will be useful for diagnostic IgE antibody assays, as well as molecular and cellular studies to investigate the immunological mechanisms of peanut allergy.

Parvalbumin In Fish Skin–Derived Gelatin: Is There A Risk For Fish Allergic Consumers?, S. J. Koppelman, J. A. Nordlee, P.-W. Lee, R. P. Happe, M. Hessing, R. Norland, T. Manning, R. Deschene, G. A. H. De Jong, S. L. Taylor

Department of Food Science and Technology: Faculty Publications

The major allergen parvalbumin was purified from cod muscle tissues, and polyclonal antibodies were raised toward it. The antibodies were tested for specificity, and an enzyme-linked immunosorbent assay (ELISA) was developed using these antibodies. The ELISA was applied to measure parvalbumin in cod skin, the starting material for fish gelatin made from deep sea, wild fish. The ELISA was sufficiently sensitive (LLOQ = 0.8 ng ml^–1 in extracts, corresponding to 0.02 μg of parvalbumin per g of tissue) and did not cross-react with common food constituents. Fish gelatin, wine, and beer, matrices for the potential use of this ELISA, …

Characterization Of Igg And Ige Binding To Parvalbumin Derived From Commercially Important Fish Species, Poi-Wah Lee

Department of Food Science and Technology: Dissertations, Theses, and Student Research

Parvalbumin is a calcium-binding muscle protein that is present in all vertebrates. Despite being a pan-allergen in fish and frog, fish-specific IgE and antiparvalbumin IgG antibodies displayed varying cross-reactivity among fish species. In this research, the parvalbumin-binding characteristics of several antibodies were investigated, including 3 IgG antibodies against frog, carp, and cod parvalbumins, and human IgE.

By immunoassay and IgG-immunoblotting, 3 antiparvalbumin antibodies revealed inconsistent specificity among 29 raw fish muscle extracts, which may be partially attributed to the decreased levels of fish muscle parvalbumin from anterior to posterior positions. Parvalbumin-binding by antibodies was unaffected in 112 days of frozen-stored …

Purification Of Parvalbumin From Carp: A Protocol That Avoids Heat Treatment, Stef J. Koppelman, Roland A. Romijn, Harmen H. J. De Jongh, Julie A. Nordlee, Sander Piersma, Martin Hessing, Steve L. Taylor

Department of Food Science and Technology: Faculty Publications

Parvalbumin from carp, a major allergen,was purified to homogeneity using ion exchange chromatography and size exclusion chromatography (estimated purity > 95% to 98% based on SDS-PAGE and native PAGE) with a yield of 318 mg, and a number of basic biochemical characteristics were determined. The identity was confirmed by peptide-mass fingerprinting, and IgE-binding was demonstrated. The UV/Vis absorbance spectra were explained using the previously published amino acid sequences. Far UV-CD spectroscopy was used to confirm the folding character of parvalbumin. We conclude that parvalbumin from carp can be purified on a comparatively large (hundreds of milligrams) scale using a purification protocol …

Practical And Predictive Bioinformatics Methods For The Identification Of Potentially Cross-Reactive Protein Matches, Richard E. Goodman

Department of Food Science and Technology: Faculty Publications

A bioinformatics comparison of proteins introduced into food crops through genetic engineering provides a mechanism to identify those proteins that may present an increased risk of allergic reactions for individuals with existing allergies. The goal is to identify proteins that are known to be allergens or are so similar to an allergen that they may induce allergic cross-reactions. Three comparative approaches have traditionally been used, or considered for safety evaluations. One identifies any short (6–8) amino acid segment of the protein that exactly matches a known allergen sequence. The second is an overall primary sequence comparison using Basic Local Alignment …