{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T18:04:41Z","timestamp":1776708281397,"version":"3.51.2"},"reference-count":56,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2022,11,3]],"date-time":"2022-11-03T00:00:00Z","timestamp":1667433600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["UIDB\/04326\/2020"],"award-info":[{"award-number":["UIDB\/04326\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["UIDP\/04326\/2020"],"award-info":[{"award-number":["UIDP\/04326\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["LA\/P\/0101\/2020"],"award-info":[{"award-number":["LA\/P\/0101\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["16-02-01-FMP-0014"],"award-info":[{"award-number":["16-02-01-FMP-0014"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"name":"ALLYFISH","award":["UIDB\/04326\/2020"],"award-info":[{"award-number":["UIDB\/04326\/2020"]}]},{"name":"ALLYFISH","award":["UIDP\/04326\/2020"],"award-info":[{"award-number":["UIDP\/04326\/2020"]}]},{"name":"ALLYFISH","award":["LA\/P\/0101\/2020"],"award-info":[{"award-number":["LA\/P\/0101\/2020"]}]},{"name":"ALLYFISH","award":["16-02-01-FMP-0014"],"award-info":[{"award-number":["16-02-01-FMP-0014"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Consumption of aquatic food, including fish, accounts for 17% of animal protein intake. However, fish consumption might also result in several side-effects such as sneezing, swelling and anaphylaxis in sensitized consumers. Fish allergy is an immune reaction to allergenic proteins in the fish muscle, for instance parvalbumin (PV), considered the major fish allergen. In this study, we characterize PV in two economically important fish species for southern European aquaculture, namely gilthead seabream and European seabass, to understand its stability during in vitro digestion and fish processing. This information is crucial for future studies on the allergenicity of processed fish products. PVs were extracted from fish muscles, identified by mass spectrometry (MS), and detected by sandwich enzyme-linked immunosorbent assay (ELISA) after simulated digestion and various food processing treatments. Secondary structures were determined by circular dichroism (CD) after purification by anion exchange and gel filtration chromatography. In both species, PVs presented as \u03b1-helical and \u03b2-sheet structures, at room temperature, were shown to unfold at boiling temperatures. In European seabass, PV detectability decreased during the simulated digestion and after 240 min (intestinal phase) no detection was observed, while steaming showed a decrease (p &lt; 0.05) in PVs detectability in comparison to raw muscle samples, for both species. Additionally, freezing (\u221220 \u00b0C) for up to 12 months continued to reduce the detectability of PV in tested processing techniques. We concluded that PVs from both species are susceptible to digestion and processing techniques such as steaming and freezing. Our study obtained preliminary results for further research on the allergenic potential of PV after digestion and processing.<\/jats:p>","DOI":"10.3390\/ani12213022","type":"journal-article","created":{"date-parts":[[2022,11,4]],"date-time":"2022-11-04T03:28:10Z","timestamp":1667532490000},"page":"3022","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Fish Processing and Digestion Affect Parvalbumins Detectability in Gilthead Seabream and European Seabass"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0274-5957","authenticated-orcid":false,"given":"Denise","family":"Schrama","sequence":"first","affiliation":[{"name":"Centre of Marine Sciences (CCMAR), Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"},{"name":"Departamento de Ci\u00eancias do Mar, da Terra e do Ambiente, Faculdade de Ci\u00eancias e Tecnologia, Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7751-2870","authenticated-orcid":false,"given":"Cl\u00e1udia","family":"Raposo de Magalh\u00e3es","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR), Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"},{"name":"Departamento de Ci\u00eancias do Mar, da Terra e do Ambiente, Faculdade de Ci\u00eancias e Tecnologia, Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7237-5053","authenticated-orcid":false,"given":"Marco","family":"Cerqueira","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR), Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"}]},{"given":"Raquel","family":"Carrilho","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR), Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"},{"name":"Departamento de Ci\u00eancias do Mar, da Terra e do Ambiente, Faculdade de Ci\u00eancias e Tecnologia, Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"}]},{"given":"Dominique","family":"Revets","sequence":"additional","affiliation":[{"name":"Department of Infection and Immunity, Luxembourg Institute of Health, 29, Rue Henri Koch, L-4354 Esch-sur-Alzette, Luxembourg"}]},{"given":"Annette","family":"Kuehn","sequence":"additional","affiliation":[{"name":"Department of Infection and Immunity, Luxembourg Institute of Health, 29, Rue Henri Koch, L-4354 Esch-sur-Alzette, Luxembourg"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5244-5541","authenticated-orcid":false,"given":"Sofia","family":"Engrola","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR), Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9668-1204","authenticated-orcid":false,"given":"Pedro M.","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR), Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"},{"name":"Departamento de Qu\u00edmica e Farm\u00e1cia, Faculdade de Ci\u00eancias e Tecnologia, Campus de Gambelas, Universidade do Algarve, 8005-139 Faro, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,3]]},"reference":[{"key":"ref_1","first-page":"427","article-title":"A review on role of fish in human nutrition with special emphasis to essential fatty acid","volume":"6","author":"Pal","year":"2018","journal-title":"Int. J. Fish. Aquat. Stud."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"103782","DOI":"10.1016\/j.jprot.2020.103782","article-title":"How tryptophan levels in plant-based aquafeeds affect fish physiology, metabolism and proteome","volume":"221","author":"Cerqueira","year":"2020","journal-title":"J. Proteom."},{"key":"ref_3","unstructured":"FAO (2022). The State of World Fisheries and Aquaculture 2022. Towards Blue Transformation, Food and Agriculture Organization."},{"key":"ref_4","unstructured":"European Commission (2021). The EU fish market: 2021 edition, European Market Observatory for Fisheries and Aquaculture Products (EUMOFA)."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Shackelford, T.K., and Weekes-Shackelford, V.A. (2021). Allergies. Encyclopedia of Evolutionary Psychological Science, Springer International Publishing.","DOI":"10.1007\/978-3-319-19650-3"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3147","DOI":"10.1016\/j.jaip.2021.04.007","article-title":"Natural History of IgE-Mediated Fish Allergy in Children","volume":"9","author":"Xepapadaki","year":"2021","journal-title":"J. Allergy Clin. Immunol. Pract."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fphys.2022.897168","article-title":"Fish Allergenicity Modulation Using Tailored Enriched Diets\u2014Where Are We?","volume":"13","author":"Schrama","year":"2022","journal-title":"Front. Physiol."},{"key":"ref_8","first-page":"2198","article-title":"Identification of Potentially Tolerated Fish Species by Multiplex IgE Testing of a Multinational Fish-Allergic Patient Cohort","volume":"22","author":"Kalic","year":"2022","journal-title":"J. Allergy Clin. Immunol. Pract."},{"key":"ref_9","unstructured":"(2022, October 01). WHO\/IUIS Allergen Nomenclature Database. Available online: http:\/\/www.allergen.org."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"414","DOI":"10.18176\/jiaci.0381","article-title":"What we know about fish allergy by the end of the decade?","volume":"29","author":"Kourani","year":"2020","journal-title":"J. Investig. Allergol. Clin. Immunol."},{"key":"ref_11","first-page":"82","article-title":"Changes in the soluble bone proteome of reared white seabream (Diplodus sargus) with skeletal deformities","volume":"6","author":"Silva","year":"2011","journal-title":"Comp. Biochem. Physiol. Part D Genom. Proteom."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1007\/s00018-008-8564-6","article-title":"The continuing disappearance of \u201cpure\u201d Ca2+ buffers","volume":"66","author":"Schwaller","year":"2009","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5577","DOI":"10.1021\/jf060291g","article-title":"Detecting Fish Parvalbumin with Commercial Mouse Monoclonal Anti-frog Parvalbumin IgG","volume":"54","author":"Chen","year":"2006","journal-title":"J. Agric. Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1159\/000316346","article-title":"Important Variations in Parvalbumin Content in Common Fish Species: A Factor Possibly Contributing to Variable Allergenicity","volume":"153","author":"Kuehn","year":"2010","journal-title":"Int. Arch. Allergy Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3211","DOI":"10.1016\/j.jprot.2012.03.030","article-title":"Rapid direct detection of the major fish allergen, parvalbumin, by selected MS\/MS ion monitoring mass spectrometry","volume":"75","author":"Carrera","year":"2012","journal-title":"J. Proteom."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.jprot.2019.04.005","article-title":"Molecular characterization of B-cell epitopes for the major fish allergen, parvalbumin, by shotgun proteomics, protein-based bioinformatics and IgE-reactive approaches","volume":"200","author":"Carrera","year":"2019","journal-title":"J. Proteom."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5363","DOI":"10.1021\/bi962436q","article-title":"Conformational Effects of Calcium Release from Parvalbumin:\u2009 Comparison of Computational Simulations with Spectroscopic Investigations","volume":"36","author":"Laberge","year":"1997","journal-title":"Biochemistry"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1951","DOI":"10.1080\/10408398.2020.1848791","article-title":"A review on food processing and preparation methods for altering fish allergenicity","volume":"62","author":"Dasanayaka","year":"2020","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1016\/0091-6749(92)90135-O","article-title":"Fish hypersensitivity. II: Clinical relevance of altered fish allergenicity caused by various preparation methods","volume":"90","author":"Strause","year":"1992","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"P\u00e9rez-Tavarez, R., Carrera, M., Pedrosa, M., Quirce, S., Rodr\u00edguez-P\u00e9rez, R., and Gasset, M. (2019). Reconstruction of fish allergenicity from the content and structural traits of the component \u03b2-parvalbumin isoforms. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-52801-6"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.jfca.2017.01.008","article-title":"A procedure for grouping food consumption data for use in food allergen risk assessment","volume":"59","author":"Birot","year":"2017","journal-title":"J. Food Compos. Anal."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Crevel, R. (2015). Food allergen risk assessment and management. Handbook of Food Allergen Detection and Control, Woodhead Publishing.","DOI":"10.1533\/9781782420217.1.41"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.biopha.2006.10.005","article-title":"Gastrointestinal digestion of food allergens: Effect on their allergenicity","volume":"61","author":"Moreno","year":"2007","journal-title":"Biomed. Pharmacother."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2000401","DOI":"10.1002\/mnfr.202000401","article-title":"Gastrointestinal Digestion of Food Proteins under the Effects of Released Bioactive Peptides on Digestive Health","volume":"64","year":"2020","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2077","DOI":"10.1080\/10408398.2015.1045965","article-title":"Review of conventional and novel food processing methods on food allergens","volume":"57","author":"Vanga","year":"2017","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"5903","DOI":"10.1021\/bi300403h","article-title":"Pressure-temperature stability, Ca2+ binding, and pressure-temperature phase diagram of cod parvalbumin: Gad m 1","volume":"51","author":"Somkuti","year":"2012","journal-title":"Biochemistry"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1159\/000242360","article-title":"Effects of Industrial Processing on the Immunogenicity of Commonly Ingested Fish Species","volume":"151","author":"Sletten","year":"2010","journal-title":"Int. Arch. Allergy Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Schrama, D., Raposo de Magalh\u00e3es, C., Cerqueira, M., Carrilho, R., Farinha, A.P., Rosa da Costa, A.M., Gon\u00e7alves, A., Kuehn, A., Revets, D., and Planchon, S. (2022). Effect of creatine and EDTA supplemented diets on European seabass (Dicentrarchus labrax) allergenicity, fish muscle quality and omics fingerprint. Comp. Biochem. Physiol. Part D Genom. Proteom., 41.","DOI":"10.1016\/j.cbd.2021.100941"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"De Magalhaes, C.R., Schrama, D., Fonseca, F., Kuehn, A., Morisset, M., Ferreira, S.R., Goncalves, A., and Rodrigues, P.M. (2020). Effect of EDTA enriched diets on farmed fish allergenicity and muscle quality; a proteomics approach. Food Chem., 305.","DOI":"10.1016\/j.foodchem.2019.125508"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Schrama, D., Cerqueira, M., Raposo, C.S., Rosa da Costa, A.M., Wulff, T., Gon\u00e7alves, A., Camacho, C., Colen, R., Fonseca, F., and Rodrigues, P.M. (2018). Dietary Creatine Supplementation in Gilthead Seabream (Sparus aurata): Comparative Proteomics Analysis on Fish Allergens, Muscle Quality, and Liver. Front. Physiol., 9.","DOI":"10.3389\/fphys.2018.01844"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1016\/j.jaip.2018.11.011","article-title":"Patients allergic to fish tolerate ray based on the low allergenicity of its parvalbumin","volume":"7","author":"Kalic","year":"2019","journal-title":"J. Allergy Clin. Immunol. Pract."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1772","DOI":"10.1111\/all.12968","article-title":"Cross-reactivity to fish and chicken meat\u2013a new clinical syndrome","volume":"71","author":"Kuehn","year":"2016","journal-title":"Allergy"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Raith, M., Zach, D., Sonnleitner, L., Woroszylo, K., Focke-Tejkl, M., Wank, H., Graf, T., Kuehn, A., Pascal, M., and Mu\u00f1oz-Cano, R.M. (2019). Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-44208-0"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1006\/abio.2000.4880","article-title":"Estimation of protein secondary structure from circular dichroism spectra: Comparison of CONTIN, SELCON, and CDSSTR methods with an expanded reference set","volume":"287","author":"Sreerama","year":"2000","journal-title":"Anal. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1039\/C3FO60702J","article-title":"A standardised static in vitro digestion method suitable for food-an international consensus","volume":"5","author":"Minekus","year":"2014","journal-title":"Food Funct."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1186\/s13601-018-0216-9","article-title":"Protease resistance of food proteins: A mixed picture for predicting allergenicity but a useful tool for assessing exposure","volume":"8","author":"Akkerdaas","year":"2018","journal-title":"Clin. Transl. Allergy"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"120","DOI":"10.5414\/ALX01485E","article-title":"Protein and DNA-based assays as complementary tools for fish allergen detection","volume":"1","author":"Kuehn","year":"2017","journal-title":"Allergol. Sel."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1016\/j.foodchem.2005.03.043","article-title":"Effects of brine concentration on shelf-life of hot-smoked tilapia (Oreochromis niloticus) stored at 4 C","volume":"97","author":"Yanar","year":"2006","journal-title":"Food Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1080\/00220973.1993.9943832","article-title":"Relative power of the Wilcoxon test, the Friedman test, and repeated-measures ANOVA on ranks","volume":"62","author":"Zimmerman","year":"1993","journal-title":"J. Exp. Educ."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1007\/s40521-019-00235-w","article-title":"Fish allergy management: From component-resolved diagnosis to unmet diagnostic needs","volume":"6","author":"Klueber","year":"2019","journal-title":"Curr. Treat. Options Allergy"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1097\/01.all.0000225163.06016.93","article-title":"The effect of gastric digestion on food allergy","volume":"6","author":"Untersmayr","year":"2006","journal-title":"Curr. Opin. Allergy Clin. Immunol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Jiang, X., and Rao, Q. (2021). Effect of Processing on Fish Protein Antigenicity and Allergenicity. Foods, 10.","DOI":"10.3390\/foods10050969"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"De Jongh, H.H., Robles, C.L., Timmerman, E., Nordlee, J.A., Lee, P.-W., Baumert, J.L., Hamilton, R.G., Taylor, S.L., and Koppelman, S.J. (2013). Digestibility and IgE-binding of glycosylated codfish parvalbumin. BioMed Res. Int., 2013.","DOI":"10.1155\/2013\/756789"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1136\/gut.46.5.661","article-title":"Purification, biochemical, and immunological characterisation of a major food allergen: Different immunoglobulin E recognition of the apo- and calcium-bound forms of carp parvalbumin","volume":"46","author":"Grote","year":"2000","journal-title":"Gut"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.bbapap.2005.06.005","article-title":"How to study proteins by circular dichroism","volume":"1751","author":"Kelly","year":"2005","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.foodres.2017.08.037","article-title":"Control of protein digestion under simulated gastrointestinal conditions using biopolymer microgels","volume":"100","author":"Zhang","year":"2017","journal-title":"Food Res. Int."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"991","DOI":"10.1038\/s41596-018-0119-1","article-title":"INFOGEST static in vitro simulation of gastrointestinal food digestion","volume":"14","author":"Brodkorb","year":"2019","journal-title":"Nat. Protoc."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1016\/j.jaci.2004.10.029","article-title":"The effects of gastric digestion on codfish allergenicity","volume":"115","author":"Untersmayr","year":"2005","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.jconrel.2015.10.022","article-title":"Intragastric pH and pressure profiles after intake of the high-caloric, high-fat meal as used for food effect studies","volume":"220","author":"Koziolek","year":"2015","journal-title":"J. Control. Release"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/S0378-4347(01)00110-4","article-title":"Stability of food allergens and allergenicity of processed foods","volume":"756","author":"Besler","year":"2001","journal-title":"J. Chromatogr. B Biomed. Sci. Appl."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"7773","DOI":"10.1080\/10408398.2021.1918629","article-title":"Non-thermal processing has an impact on the digestibility of the muscle proteins","volume":"62","author":"Bhat","year":"2021","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1016\/j.foodchem.2012.05.030","article-title":"Measuring parvalbumin levels in fish muscle tissue: Relevance of muscle locations and storage conditions","volume":"135","author":"Lee","year":"2012","journal-title":"Food Chem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.foodchem.2013.10.035","article-title":"Antibody reactivity to the major fish allergen parvalbumin is determined by isoforms and impact of thermal processing","volume":"148","author":"Saptarshi","year":"2014","journal-title":"Food Chem."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/j.foodchem.2015.08.037","article-title":"Quantification of major allergen parvalbumin in 22 species of fish by SDS\u2013PAGE","volume":"194","author":"Kobayashi","year":"2016","journal-title":"Food Chem."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"125333","DOI":"10.1016\/j.foodchem.2019.125333","article-title":"Lipid emulsion enhances fish allergen parvalbumin\u2019s resistance to in vitro digestion and IgG\/IgE binding capacity","volume":"302","author":"Luo","year":"2020","journal-title":"Food Chem."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1016\/j.foodchem.2012.08.066","article-title":"A sensitive enzyme-linked immunosorbent assay for the determination of fish protein in processed foods","volume":"136","author":"Shibahara","year":"2013","journal-title":"Food Chem."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/12\/21\/3022\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:10:05Z","timestamp":1760145005000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/12\/21\/3022"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,3]]},"references-count":56,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["ani12213022"],"URL":"https:\/\/doi.org\/10.3390\/ani12213022","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,3]]}}}