foodsystFood systemsПищевые системы2618-97712618-7272Федеральный научный центр пищевых систем им. В.М. Горбатова РАН10.21323/2618-9771-2026-9-2-306-315foodsyst-1100Research ArticleСтатьиEvaluation of cheese whey protein precipitation methods based on efficiency and fractional compositionОценка методов осаждения белков подсырной сыворотки по эффективности и фракционному составуhttps://orcid.org/0000-0002-2145-1439МамыкинД. С.MamykinD. S.

Мамыкин Денис Станиславович – кандидат технических наук, научный сотрудник, отдел микробиологических исследований

152613, Ярославская область, Углич, Красноармейский бульвар, 19

Denis S. Mamykin, Candidate of Technical Sciences, Researcher, Department of Microbiological Research

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

d.mamykin@fncps.ruhttps://orcid.org/0000-0002-7665-8517ШухаловаО. М.ShukhalovaO. М.

Шухалова Ольга Михайловна – кандидат технических наук, руководитель, отдел микробиологических исследований

152613, Ярославская область, Углич, Красноармейский бульвар, 19

Olga М. Shukhalova, Candidate of Technical Sciences, Supervisor, Department of Microbiological Research

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

o.shukhalova@fncps.ruhttps://orcid.org/0000-0002-1863-009XВолковаТ. А.VolkovaT. A.

Волкова Татьяна Алексеевна – кандидат технических наук, ученый секретарь

152613, Ярославская область, Углич, Красноармейский бульвар, 19

Tatiana A. Volkova, Candidate of Technical Sciences, scientific secretary

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

t.volkova@fncps.ruhttps://orcid.org/0009-0001-7948-8840БородинаУ. В.BorodinaU. V.

Бородина Ульяна Владимировна – инженер I категории, отдел микробиологических исследований

152613, Ярославская область, Углич, Красноармейский бульвар, 19

Ulyana V. Borodina, 1st Category Engineer, Department of Microbiological Research

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

u.borodina@fncps.ruhttps://orcid.org/0000-0003-4364-0342ГригорьеваА. И.GrigorievaA. I.

Григорьева Анастасия Игоревна – младший научный сотрудник, отдел физико-химических и биохимических исследований

152613, Ярославская область, Углич, Красноармейский бульвар, 19

Anastasia I. Grigorieva, Junior Researcher, Department of Physical, Chemical and Biochemical Research

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

a.grigoriyeva@fncps.ruВсероссийский научно-исследовательский институт маслоделия и сыроделияРоссияAll-Russian Scientific Research Institute of Butter- and CheesemakingRussian Federation20260108202692306315Copyright © Mamykin D.S., Shukhalova O.М., Volkova T.A., Borodina U.V., Grigorieva A.I., 20262026Мамыкин Д.С., Шухалова О.М., Волкова Т.А., Бородина У.В., Григорьева А.И.Mamykin D.S., Shukhalova O.М., Volkova T.A., Borodina U.V., Grigorieva A.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.fsjour.com/jour/article/view/1100

Cheese whey is a large-scale by-product of milk processing, characterized by high biological value. However, its irrational use can lead to a serious environmental burden. One of the ways to solve this problem is the isolation of proteins and their fractionation, the efficiency of which significantly depends on the method used. In this work, a comparative assessment of four methods of processing reconstituted cheese whey (6.0 % dry matter mass fraction) was carried out, such as chitosan precipitation (pH 4.5; 20 °C; 1 % gel; 20 min), thermal acid coagulation (pH 4.65; 90 °C; 15 min) using lactic, citric, phosphoric and hydrochloric acids, cryo-acid precipitation (pH 4.5; –18 °C; 24 h) and ultrafiltration (spiral membranes with a cutoff threshold of 10 kDa). The study determined the proportion of precipitated protein, the fractional composition of soluble nitrogenous compounds using gel filtration, and specific operating costs. Ultrafiltration provided the highest efficiency of protein separation (65.92 ± 0.59 %), which also turned out to be an order of magnitude cheaper (≈112 rubles/m3) than all chemical methods. Among the thermal acid options, citric (57.83 ± 0.51 %) and hydrochloric (56.63 ± 0.55 %) acids proved to be the most effective. Chitosan and cryo-acid treatment showed low overall efficiency (26.51 ± 0.20 % and 25.30 ± 0.15 %, respectively). Analysis of the fractional composition of the supernatant showed that thermal acid coagulation and ultrafiltration completely removed β-lactoglobulin and BSA. Chitosan precipitation preserved the native profile of all major whey proteins, removing only ballast impurities. The cryoacid method demonstrated unique selectivity, virtually completely removing peptides while preserving native α-lactalbumin and β-lactoglobulin. A step-by-step fractionation strategy is proposed: treatment with chitosan to remove impurities, cryoacid separation of peptides, followed by isolation of native proteins by ultrafiltration or the thermal acid method.

Подсырная сыворотка является крупнотоннажным побочным продуктом переработки молока и характеризуется высокой биологической ценностью. Однако ее нерациональное использование может приводить к существенной экологической нагрузке. Одним из способов решения данной проблемы является выделение и фракционирование белков, эффективность которых существенно зависит от применяемого метода. В работе проведена сравнительная оценка четырех способов обработки восстановленной подсырной сыворотки (массовая доля сухих веществ 6,0 %), таких как осаждение хитозаном (pH 4,5; 20 °C; 1 % геля; 20 мин), термокислотная коагуляция (pH 4,65; 90 °C; 15 мин) с использованием молочной, лимонной, ортофосфорной и соляной кислот, криокислотное осаждение (pH 4,5; минус 18 °C; 24 ч) и ультрафильтрация (спиральные мембраны с порогом отсечения 10 кДа). В ходе исследования определяли долю выделенного белка, фракционный состав растворимых азотистых соединений методом гель-фильтрации и удельные операционные затраты. Максимальную эффективность отделения белка обеспечила ультрафильтрация (65,92 ± 0,59 %), которая оказалась на порядок дешевле (≈ 112 руб/м3) всех химических методов. Среди термокислотных вариантов наиболее эффективными показали себя лимонная (57,83 ± 0,51 %) и соляная (56,63 ± 0,55 %) кислоты. Хитозан и крио-кислотная обработка показали низкую общую эффективность (26,51 ± 0,20 % и 25,30 ± 0,15 % соответственно). Анализ фракционного состава супернатанта продемонстрировал, что термокислотная коагуляция и ультрафильтрация привели к полному удалению β-лактоглобулина и БСА. Осаждение хитозаном обеспечило сохранение нативного профиля всех основных сывороточных белков и способствовало удалению лишь балластных примесей. Криокислотный метод продемонстрировал уникальную селективность, практически полное удаление пептидов при сохранении нативных α-лактальбумина и β-лактоглобулина. Предложена поэтапная стратегия фракционирования: обработка хитозаном для удаления примесей, отделение пептидов криокислотным способом с последующим выделением нативных белков ультрафильтрацией или термокислотным методом.

сывороточные белкихитозанденатурациякислотно-щелочная коагуляцияизоэлектрическое осаждениеwhey proteinschitosandenaturationacid-base coagulationisoelectric precipitationCтатья подготовлена в рамках выполнения исследований по государственному заданию № FGUS‑2024-0007 Федерального научного центра пищевых систем им. В. М. Горбатова Российской академии наук.The article was prepared as part of research carried out on a government assignment № FGUS‑2024–0007 V. M. Gorbatov Federal Research Center for Food Systems of the Russian Academy of Sciences.ReferencesVanany, I., Wangsa, I. D., Jeremi, N. A. (2024). A multi-objective mixed-integer linear model for sustainable dairy supply chain with food waste and environmental pollutants. Process Integration and Optimization for Sustainability, 8, 723–740. https://doi.org/10.1007/s41660-023-00382-3.Vanany, I., Wangsa, I. D., Jeremi, N. A. (2024). A multi-objective mixed-integer linear model for sustainable dairy supply chain with food waste and environmental pollutants. Process Integration and Optimization for Sustainability, 8, 723–740. https://doi.org/10.1007/s41660-023-00382-3.Buchanan, D., Martindale, W., Romeih, E., Hebishy, E. (2023). Recent advances in whey processing and valorisation: Technological and environmental perspectives. International Journal of Dairy Technology, 76(2), 291–312. https://doi.org/10.1111/1471-0307.12935.Buchanan, D., Martindale, W., Romeih, E., Hebishy, E. (2023). Recent advances in whey processing and valorisation: Technological and environmental perspectives. International Journal of Dairy Technology, 76(2), 291–312. https://doi.org/10.1111/1471-0307.12935.Mota, J., Moura-Alves, M., Teixeira, A. F., Nóbrega, R., Lameirão, D., Gonçalves, C. (2026). From cheese whey to functional ingredients: Upcycling whey proteins for cardiovascular and immunomodulatory health — evidence mapping and perspectives from Portugal. Foods, 15(5), Article 908. https://doi.org/10.3390/foods15050908.Mota, J., Moura-Alves, M., Teixeira, A. F., Nóbrega, R., Lameirão, D., Gonçalves, C. (2026). From cheese whey to functional ingredients: Upcycling whey proteins for cardiovascular and immunomodulatory health — evidence mapping and perspectives from Portugal. Foods, 15(5), Article 908. https://doi.org/10.3390/foods15050908.Sukmana, H., Rizwana, I., Dobozi, R., Al-Tayawi, A. N., Szabó, B. P., Özer, B. et al. (2025). A review of various protein separation techniques and valorization of cheese whey and buttermilk. European Food Research and Technology, 251(12), 4263–4279. https://doi.org/10.1007/s00217-025-04877-w.Sukmana, H., Rizwana, I., Dobozi, R., Al-Tayawi, A. N., Szabó, B. P., Özer, B. et al. (2025). A review of various protein separation techniques and valorization of cheese whey and buttermilk. European Food Research and Technology, 251(12), 4263–4279. https://doi.org/10.1007/s00217-025-04877-w.Костенко, К. В., Костенко, Е. Г., Пойдун, Ф. А., Рехман, З. А., Аскерова, А. С., Голик, Д. Б. и др. (2026). Оптимизация процесса восстановления вторичных молочных продуктов с применением ультразвуковой обработки и исследование их стабильности. Аграрный вестник Урала, 26(1), 140–151.[Kostenko, K. V., Kostenko, E. G., Poydun, F. A., Rekhman, Z. A., Askerova, A. S., Golik, D. B. et al. (2026). Optimization of the regeneration process for secondary dairy products using ultrasonic treatment and a study of their stability. Agrarian Bulletin of the Urals, 26(1), 140–151. (In Russian)] https://doi.org/10.32417/1997-4868-2026-26-01-140-151.Свириденко, Ю. Я., Кравченко, Е. Ф., Яковлева, О. А. (2008). Использование молочной сыворотки и локальная очистка сточных вод. Молочная промышленность, 11, 58–60.[Sviredenko, Yu. Ya., Kravchenko, E. F., Yakovleva, O. A. (2008). Milk whey application and local purification of effluents. Dairy Industry, 11, 58–60. (In Russian)].Semproli, R., Cassano, L., Ballabio, G., Cappelletti, G., Speranza, G., Donzella, S. et al. (2025). Upcycling cheese whey permeate into fully bio-based surfactants through fermentation and biocatalysis. Applied Microbiology and Biotechnology, 109(1), Article 270. https://doi.org/10.1007/s00253-025-13630-y.Semproli, R., Cassano, L., Ballabio, G., Cappelletti, G., Speranza, G., Donzella, S. et al. (2025). Upcycling cheese whey permeate into fully bio-based surfactants through fermentation and biocatalysis. Applied Microbiology and Biotechnology, 109(1), Article 270. https://doi.org/10.1007/s00253-025-13630-y.Usmani, Z., Sharma, M., Gaffey, J., Sharma, M., Dewhurst, R. J., Moreau, B. et al. (2022). Valorization of dairy waste and by-products through microbial bioprocesses. Bioresource Technology, 346, Article 126444. https://doi.org/10.1016/j.biortech.2021.126444.Usmani, Z., Sharma, M., Gaffey, J., Sharma, M., Dewhurst, R. J., Moreau, B. et al. (2022). Valorization of dairy waste and by-products through microbial bioprocesses. Bioresource Technology, 346, Article 126444. https://doi.org/10.1016/j.biortech.2021.126444.Mirzakulova, A., Sarsembaeva, T., Suleimenova, Z., Kowalski, Ł., Gajdzik, B., Wolniak, R. et al. (2025). Whey: Composition, processing, application, and prospects in functional and nutritional beverages — A review. Foods, 14(18), Article 3245. https://doi.org/10.3390/foods14183245.Mirzakulova, A., Sarsembaeva, T., Suleimenova, Z., Kowalski, Ł., Gajdzik, B., Wolniak, R. et al. (2025). Whey: Composition, processing, application, and prospects in functional and nutritional beverages — A review. Foods, 14(18), Article 3245. https://doi.org/10.3390/foods14183245.Tsermoula, P., Khakimov, B., Nielsen, J. H., Engelsen, S. B. (2021). WHEY-The wastestream that became more valuable than the food product. Trends in Food Science and Technology, 118(Part A), 230–241. https://doi.org/10.1016/j.tifs.2021.08.025.Tsermoula, P., Khakimov, B., Nielsen, J. H., Engelsen, S. B. (2021). WHEY-The wastestream that became more valuable than the food product. Trends in Food Science and Technology, 118(Part A), 230–241. https://doi.org/10.1016/j.tifs.2021.08.025.Juthi, L. M., Kabir, T., Podder, N., Bhuiyan, M. H. R., Yeasmen, N., Aziz, M. G. (2025). Separation and characterization of whey protein powder from cheese byproduct. Applied Food Research, 5(2), Article 101398. https://doi.org/10.1016/j.afres.2025.101398.Juthi, L. M., Kabir, T., Podder, N., Bhuiyan, M. H. R., Yeasmen, N., Aziz, M. G. (2025). Separation and characterization of whey protein powder from cheese byproduct. Applied Food Research, 5(2), Article 101398. https://doi.org/10.1016/j.afres.2025.101398.Kumar, L. R. G., Mohanan, V., Pavithra, P. A., Anas, K. K. (2026). Encapsulation of squalene in oil-in-water emulsions: Utilization of heat-polymerized whey proteins as emulsifiers. International Journal of Biological Macromolecules, 352 Article 151090. https://doi.org/10.1016/j.ijbiomac.2026.151090.Kumar, L. R. G., Mohanan, V., Pavithra, P. A., Anas, K. K. (2026). Encapsulation of squalene in oil-in-water emulsions: Utilization of heat-polymerized whey proteins as emulsifiers. International Journal of Biological Macromolecules, 352 Article 151090. https://doi.org/10.1016/j.ijbiomac.2026.151090.Chawla, R., Roy, S., Malik, B. (2023). Valorisation of Whey for Development of Different Types of Food Products Including Fermented Beverages. Chapter in a book: Whey Valorization. Springer, Singapore, 2023. https://doi.org/10.1007/978-981-99-5459-9_11.Chawla, R., Roy, S., Malik, B. (2023). Valorisation of Whey for Development of Different Types of Food Products Including Fermented Beverages. Chapter in a book: Whey Valorization. Springer, Singapore, 2023. https://doi.org/10.1007/978-981-99-5459-9_11.Chen, G. Q., Qu, Y., Gras, S. L., Kentish, S. E. (2023). Separation technologies for whey protein fractionation. Food Engineering Reviews, 15(3), 438–465. https://doi.org/10.1007/s12393-022-09330-2.Chen, G. Q., Qu, Y., Gras, S. L., Kentish, S. E. (2023). Separation technologies for whey protein fractionation. Food Engineering Reviews, 15(3), 438–465. https://doi.org/10.1007/s12393-022-09330-2.Prabhakaran, G. S., Molitor, M., Govindasamy-Lucey, S., Lucey, J. A. (2024). Heat-stable whey protein isolate made using isoelectric precipitation and clarification. Journal of Dairy Science, 107(9), 6629–6642. https://doi.org/10.3168/jds.2024-24671.Prabhakaran, G. S., Molitor, M., Govindasamy-Lucey, S., Lucey, J. A. (2024). Heat-stable whey protein isolate made using isoelectric precipitation and clarification. Journal of Dairy Science, 107(9), 6629–6642. https://doi.org/10.3168/jds.2024-24671.Lamkaddam, I. U., Vega, E., Colón, J., Ponsá, S., Llenas, L., Mora, M. (2023). Progressive freeze concentration of cheese whey for protein and lactose recovery. International Dairy Journal, 139, Article 105572. https://doi.org/10.1016/j.idairyj.2022.105572.Lamkaddam, I. U., Vega, E., Colón, J., Ponsá, S., Llenas, L., Mora, M. (2023). Progressive freeze concentration of cheese whey for protein and lactose recovery. International Dairy Journal, 139, Article 105572. https://doi.org/10.1016/j.idairyj.2022.105572.Al Kader, H., Gill, H., Truong, T. (2023). Demineralisation and recovery of whey proteins from commercial full-fat salty Cheddar whey using size-exclusion chromatography. Food Chemistry, 405(1–3), Article 134831. https://doi.org/10.1016/j.foodchem.2022.134831.Al Kader, H., Gill, H., Truong, T. (2023). Demineralisation and recovery of whey proteins from commercial full-fat salty Cheddar whey using size-exclusion chromatography. Food Chemistry, 405(1–3), Article 134831. https://doi.org/10.1016/j.foodchem.2022.134831.Pellegrino, L., Hogenboom, J. A., Rosi, V., Sindaco, M., Gerna, S., D’Incecco, P. (2022). Focus on the protein fraction of sports nutrition supplements. Molecules, 27(11), Article 3487. https://doi.org/10.3390/molecules27113487.Pellegrino, L., Hogenboom, J. A., Rosi, V., Sindaco, M., Gerna, S., D’Incecco, P. (2022). Focus on the protein fraction of sports nutrition supplements. Molecules, 27(11), Article 3487. https://doi.org/10.3390/molecules27113487.Patel, S. (2015). Functional food relevance of whey protein: A review of recent findings and scopes ahead. Journal of Functional Foods, 19(10), 308–319. https://doi.org/10.1016/j.jff.2015.09.040Patel, S. (2015). Functional food relevance of whey protein: A review of recent findings and scopes ahead. Journal of Functional Foods, 19(10), 308–319. https://doi.org/10.1016/j.jff.2015.09.040Cereda, E., Pisati, R., Rondanelli, M., Caccialanza, R. (2022). Whey protein, leucine- and vitamin-D-enriched oral nutritional supplementation for the treatment of sarcopenia. Nutrients, 14(7), Article 1524. https://doi.org/10.3390/nu14071524.Cereda, E., Pisati, R., Rondanelli, M., Caccialanza, R. (2022). Whey protein, leucine- and vitamin-D-enriched oral nutritional supplementation for the treatment of sarcopenia. Nutrients, 14(7), Article 1524. https://doi.org/10.3390/nu14071524.Ryan, M. P., Walsh, G. (2016). The biotechnological potential of whey. Reviews in Environmental Science and Biotechnology, 15(3), 479–498. https://doi.org/10.1007/s11157-016-9402-1Ryan, M. P., Walsh, G. (2016). The biotechnological potential of whey. Reviews in Environmental Science and Biotechnology, 15(3), 479–498. https://doi.org/10.1007/s11157-016-9402-1Uribarrena, M., Rovira-Cal, E., Urbina, L., Suárez, M. J., Aymerich, E., Guerrero, P. et al. (2024). Valorization of cheese whey: Closing the loop from protein extraction to whey protein film composting. Green Chemistry, 26(7), 4103–4111. https://doi.org/10.1039/D3GC04304E.Uribarrena, M., Rovira-Cal, E., Urbina, L., Suárez, M. J., Aymerich, E., Guerrero, P. et al. (2024). Valorization of cheese whey: Closing the loop from protein extraction to whey protein film composting. Green Chemistry, 26(7), 4103–4111. https://doi.org/10.1039/D3GC04304E.Гаврилов, Г. Б., Гаврилов, Б. Г. (2009). Закономерности мембранного кон- центрирования сывороточных белков. Техника и технология пищевых производств, 1(12), 26–29.[Gavrilov, G. B., Gavrilov, B. G. (2009). Laws membran concentration whey fibers. Food Processing: Techniques and Technology, 1(12), 26–29. (In Russian)].Храмцов, А. Г. (2021). Технологический прорыв агропищевых инноваций молочного дела на примере универсального сельхозсырья. Обратный осмос. Аграрно-пищевые инновации, 2(14), 7–20.[Khramtsov, A. G. (2021). Technological breakthrough of agrarian-and-food innovations in dairy case for example of universal agricultural raw materials. Reverse osmosis. Agrarian-and-Food Innovations, 2(14), 7–20. (In Russian)] https://doi.org/10.31208/2618-7353-2021-14-7-20.Abish, Z. A., Alibekov, R. S., Tarapoulouzi, M., Bakhtybekova, A. R., Kobzhasarova, Z. I. (2024). Review in deep processing of whey. Cogent Food and Agriculture, 10(1), Article 2415380. https://doi.org/10.1080/23311932.2024.2415380.Abish, Z. A., Alibekov, R. S., Tarapoulouzi, M., Bakhtybekova, A. R., Kobzhasarova, Z. I. (2024). Review in deep processing of whey. Cogent Food and Agriculture, 10(1), Article 2415380. https://doi.org/10.1080/23311932.2024.2415380.Paladii, I. V., Vrabie, E. G., Sprinchan, K. G., Bologa, M. K. (2021). Whey: Review. Part 2. Treatment processes and methods. Surface Engineering and Applied Electrochemistry, 57, 651–666. https://doi.org/10.3103/S1068375521060119.Paladii, I. V., Vrabie, E. G., Sprinchan, K. G., Bologa, M. K. (2021). Whey: Review. Part 2. Treatment processes and methods. Surface Engineering and Applied Electrochemistry, 57, 651–666. https://doi.org/10.3103/S1068375521060119.Pires, A. F., Marnotes, N. G., Rubio, O. D., Garcia, A. C., Pereira, C. D. (2021). Dairy by-products: A review on the valorization of whey and second cheese whey. Foods, 10(5), Article 1067. https://doi.org/10.3390/foods10051067.Pires, A. F., Marnotes, N. G., Rubio, O. D., Garcia, A. C., Pereira, C. D. (2021). Dairy by-products: A review on the valorization of whey and second cheese whey. Foods, 10(5), Article 1067. https://doi.org/10.3390/foods10051067.Khalid, M., Ghasem, N., Yang, X., Schroen, K., Nazir, A. (2025). Progress in membrane technology for the enrichment of individual whey proteins. Industrial and Engineering Chemistry Research, 64(40), 19305–19321. https://doi.org/10.1021/acs.iecr.5c00643.Khalid, M., Ghasem, N., Yang, X., Schroen, K., Nazir, A. (2025). Progress in membrane technology for the enrichment of individual whey proteins. Industrial and Engineering Chemistry Research, 64(40), 19305–19321. https://doi.org/10.1021/acs.iecr.5c00643.Барковская, И. А., Кручинин, А. Г. (2024). Исследование процесса селективной термоденатурации белков молочной сыворотки. Пищевая промышленность, 11, 34–37.[Barkovskaya, I. A., Kruchinin, A. G. (2024). Study of the process of selective thermodenaturation of whey proteins. Food Industry, 11, 34–37. (In Russian)] https://doi.org/10.52653/PPI.2024.11.11.006.Casas, V. E. G., Pérez, J. V., Companioni, R. S., García, N. D. (2021). Protein recovery from residual bovine whey: Influence of acid thermo-coagulation. Revista Bionatura, 6(1), 1602–1607. https://doi.org/10.21931/RB/2021.06.01.24.Casas, V. E. G., Pérez, J. V., Companioni, R. S., García, N. D. (2021). Protein recovery from residual bovine whey: Influence of acid thermo-coagulation. Revista Bionatura, 6(1), 1602–1607. https://doi.org/10.21931/RB/2021.06.01.24.Храмцов, А. Г. (2014). Логистика формирования технологической платформы получения биокластеров жира и белков из молочной сыворотки. Техника и технология пищевых производств, 2(33), 102–107.[Hramtsov, A. G. (2014). Formation logistics of technological platform for obtaining lipid and protein bio-clusters from whey. Food Processing: Techniques and Technology, 2(33), 102–107. (In Russian)].Toro-Sierra, J., Tolkach, A., Kulozik, U. (2013). Fractionation of α-lactalbumin and β-lactoglobulin from whey protein isolate using selective thermal aggregation, an optimized membrane separation procedure and resolubilization techniques at pilot plant scale. Food and Bioprocess Technology, 6(4), 1032–1043. https://doi.org/10.1007/s11947-011-0732-2.Toro-Sierra, J., Tolkach, A., Kulozik, U. (2013). Fractionation of α-lactalbumin and β-lactoglobulin from whey protein isolate using selective thermal aggregation, an optimized membrane separation procedure and resolubilization techniques at pilot plant scale. Food and Bioprocess Technology, 6(4), 1032–1043. https://doi.org/10.1007/s11947-011-0732-2.Aider, M., De Halleux, D., Akbache, A. (2007). Whey cryoconcentration and impact on its composition. Journal of Food Engineering, 82(1), 92–102. https://doi.org/10.1016/j.jfoodeng.2007.01.024.Aider, M., De Halleux, D., Akbache, A. (2007). Whey cryoconcentration and impact on its composition. Journal of Food Engineering, 82(1), 92–102. https://doi.org/10.1016/j.jfoodeng.2007.01.024.Hanušová, J., Mihulova, M., Diblikova, L., Čurda, L. (2014). Influence of salts on selective coagulation of whey proteins and their application in the isolation of β-lactoglobulin. Czech Journal of Food Sciences, 32(1), 77–81. https://doi.org/10.17221/524/2012-CJFS.Hanušová, J., Mihulova, M., Diblikova, L., Čurda, L. (2014). Influence of salts on selective coagulation of whey proteins and their application in the isolation of β-lactoglobulin. Czech Journal of Food Sciences, 32(1), 77–81. https://doi.org/10.17221/524/2012-CJFS.He, L., Ma, J., Lu, H., Li, Q., Liang, T., Shang, J. (2026). Recovery of active proteins from whey via a novel hybrid process of chitosan extraction and electrodialysis with ultrafiltration membrane. Chemical Engineering Science, 330, Article 123849. https://doi.org/10.1016/j.ces.2026.123849.He, L., Ma, J., Lu, H., Li, Q., Liang, T., Shang, J. (2026). Recovery of active proteins from whey via a novel hybrid process of chitosan extraction and electrodialysis with ultrafiltration membrane. Chemical Engineering Science, 330, Article 123849. https://doi.org/10.1016/j.ces.2026.123849.Nath, A., Zin, M. M., Molnár, M. A., Bánvölgyi, S., Gáspár, I., Vatai, G. et al. (2022). Membrane chromatography and fractionation of proteins from whey — A review. Processes, 10(5), Article 1025. https://doi.org/10.3390/pr10051025.Nath, A., Zin, M. M., Molnár, M. A., Bánvölgyi, S., Gáspár, I., Vatai, G. et al. (2022). Membrane chromatography and fractionation of proteins from whey — A review. Processes, 10(5), Article 1025. https://doi.org/10.3390/pr10051025.Romo, M., Bou, R., Castellari, M., Felipe, X. (2026). Caprine and bovine cheese whey valorization: High-pressure processing for α-lactalbumin fractionation from whey concentrates. International Dairy Journal, 175, Article 106553. https://doi.org/10.1016/j.idairyj.2026.106553.Romo, M., Bou, R., Castellari, M., Felipe, X. (2026). Caprine and bovine cheese whey valorization: High-pressure processing for α-lactalbumin fractionation from whey concentrates. International Dairy Journal, 175, Article 106553. https://doi.org/10.1016/j.idairyj.2026.106553.Rama, G. R., Timmers, L. F. S. M., de Souza, C. F. V. (2021). Ultrafiltration of cheese whey: Achieving high protein rejection and sustaining membrane efficiency. Journal of Food Processing and Preservation, 45(11), Article e15908. https://doi.org/10.1111/jfpp.15908.Rama, G. R., Timmers, L. F. S. M., de Souza, C. F. V. (2021). Ultrafiltration of cheese whey: Achieving high protein rejection and sustaining membrane efficiency. Journal of Food Processing and Preservation, 45(11), Article e15908. https://doi.org/10.1111/jfpp.15908.Wang, W. Q., Han, X., Yi, H., Zhang, L. W. (2018). The ultrafiltration efficiency and mechanism of transglutaminase enzymatic membrane reactor (EMR) for protein recovery from cheese whey. International Dairy Journal, 80, 52–61. https://doi.org/10.1016/j.idairyj.2017.12.012.Wang, W. Q., Han, X., Yi, H., Zhang, L. W. (2018). The ultrafiltration efficiency and mechanism of transglutaminase enzymatic membrane reactor (EMR) for protein recovery from cheese whey. International Dairy Journal, 80, 52–61. https://doi.org/10.1016/j.idairyj.2017.12.012.Casal, E., Montilla, A., Moreno, F. J., Olano, A., Corzo, N. (2006). Use of chitosan for selective removal of β-lactoglobulin from whey. Journal of Dairy Science, 89(5), 1384–1389. https://doi.org/10.3168/jds.S0022-0302(06)72206-8.Casal, E., Montilla, A., Moreno, F. J., Olano, A., Corzo, N. (2006). Use of chitosan for selective removal of β-lactoglobulin from whey. Journal of Dairy Science, 89(5), 1384–1389. https://doi.org/10.3168/jds.S0022-0302(06)72206-8.Cheng, Z., van Leusden, P., Eisner, M. D., Ahrné, L. (2026). Effect of the protein content and amount of native whey protein on properties of heat and acidinduced milk gels. International Dairy Journal, 179, Article 106643. https://doi.org/10.1016/j.idairyj.2026.106643.Cheng, Z., van Leusden, P., Eisner, M. D., Ahrné, L. (2026). Effect of the protein content and amount of native whey protein on properties of heat and acidinduced milk gels. International Dairy Journal, 179, Article 106643. https://doi.org/10.1016/j.idairyj.2026.106643.Anema, S. G. (2021). Heat-induced changes in caseins and casein micelles, including interactions with denatured whey proteins. International Dairy Journal, 122, Article 105136. https://doi.org/10.1016/j.idairyj.2021.105136.Anema, S. G. (2021). Heat-induced changes in caseins and casein micelles, including interactions with denatured whey proteins. International Dairy Journal, 122, Article 105136. https://doi.org/10.1016/j.idairyj.2021.105136.Kharlamova, A., Nicolai, T., Chassenieux, C. (2019). Heat-induced gelation of mixtures of casein micelles with whey protein aggregates. Food Hydrocolloids, 92(4), 198–207. https://doi.org/10.1016/j.foodhyd.2019.01.048.Kharlamova, A., Nicolai, T., Chassenieux, C. (2019). Heat-induced gelation of mixtures of casein micelles with whey protein aggregates. Food Hydrocolloids, 92(4), 198–207. https://doi.org/10.1016/j.foodhyd.2019.01.048.El-Tanboly, E. S., El-Hofi, M. K., Khorshid. (2017). Recovery of cheese whey, a by-product from the dairy industry for use as an animal feed. Journal of Nutrition and Health Food Engineering, 6(5), 148–153. https://doi.org/10.15406/jnhfe.2017.06.00215.El-Tanboly, E. S., El-Hofi, M. K., Khorshid. (2017). Recovery of cheese whey, a by-product from the dairy industry for use as an animal feed. Journal of Nutrition and Health Food Engineering, 6(5), 148–153. https://doi.org/10.15406/jnhfe.2017.06.00215.Sebastián-Nicolás, J. L., González-Olivares, L. G., Vázquez-Rodríguez, G. A., Lucho-Constatino, C. A., Castañeda-Ovando, A., Cruz-Guerrero, A. E. (2020). Valorization of whey using a biorefinery. Biofuels, Bioproducts and Biorefining, 14(5), 1010–1027. https://doi.org/10.1002/bbb.2100.Sebastián-Nicolás, J. L., González-Olivares, L. G., Vázquez-Rodríguez, G. A., Lucho-Constatino, C. A., Castañeda-Ovando, A., Cruz-Guerrero, A. E. (2020). Valorization of whey using a biorefinery. Biofuels, Bioproducts and Biorefining, 14(5), 1010–1027. https://doi.org/10.1002/bbb.2100.Souza, H. K. S., Gonçalves, M. D. P., Gómez, J. (2011). Effect of Chitosan Degradation on its Interaction with β-Lactoglobulin. Biomacromolecules, 12(4), 1015–1023, https://doi.org/10.1021/bm101356g.Souza, H. K. S., Gonçalves, M. D. P., Gómez, J. (2011). Effect of Chitosan Degradation on its Interaction with β-Lactoglobulin. Biomacromolecules, 12(4), 1015–1023, https://doi.org/10.1021/bm101356g.Jarto, I., Lucey, J. A., Molitor, M. S., Smith, K. E. (2015). Utilisation of chitosan flocculation of residual lipids and microfiltration for the production of low fat, clear WPC 80. International Journal of Dairy Technology, 68(4), 471–477. https://doi.org/10.1111/1471-0307.12249.Jarto, I., Lucey, J. A., Molitor, M. S., Smith, K. E. (2015). Utilisation of chitosan flocculation of residual lipids and microfiltration for the production of low fat, clear WPC 80. International Journal of Dairy Technology, 68(4), 471–477. https://doi.org/10.1111/1471-0307.12249.Cochachín-Guerra, B., Moreno-Cuevas, J., Carvajal-Mena, N. (2023). Effects of concentration by block freezing and vacuum evaporation on the physicochemical properties and digestibility of whey. CyTA-Journal of Food, 21(1), 313–320. https://doi.org/10.1080/19476337.2023.2196321.Cochachín-Guerra, B., Moreno-Cuevas, J., Carvajal-Mena, N. (2023). Effects of concentration by block freezing and vacuum evaporation on the physicochemical properties and digestibility of whey. CyTA-Journal of Food, 21(1), 313–320. https://doi.org/10.1080/19476337.2023.2196321.Dobozi, R., Jákói, Z. P., Beszédes, S., Szabó, B. P., Kertész, S. (2026). Investigating the separation efficiency of ultrafiltration/diafiltration (UF/DF) of whey by dielectric measurements. Sustainable Chemistry, 7(1), Article 1. https://doi.org/10.3390/suschem7010001.Dobozi, R., Jákói, Z. P., Beszédes, S., Szabó, B. P., Kertész, S. (2026). Investigating the separation efficiency of ultrafiltration/diafiltration (UF/DF) of whey by dielectric measurements. Sustainable Chemistry, 7(1), Article 1. https://doi.org/10.3390/suschem7010001.

The authors declare that there are no conflicts of interest present.