{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T13:44:04Z","timestamp":1761831844505,"version":"build-2065373602"},"reference-count":37,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T00:00:00Z","timestamp":1761782400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T00:00:00Z","timestamp":1761782400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100002641","name":"Konkuk University","doi-asserted-by":"publisher","award":["This paper was supported by Konkuk University in 2025 (S.Y.C)."],"award-info":[{"award-number":["This paper was supported by Konkuk University in 2025 (S.Y.C)."]}],"id":[{"id":"10.13039\/501100002641","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"crossref","award":["2020R1I1A3071938"],"award-info":[{"award-number":["2020R1I1A3071938"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Blinatumomab is an anti-cancer Bispecific T-cell Engager (BiTE) antibody drug approved by the Food and Drug Administration (FDA) for treating acute lymphoblastic leukemia (ALL). It targets CD3\u03b5, which is expressed on T cells and concomitantly targets CD19, an antigen specifically expressed in B-cell leukemia. Blinatumomab-bound T cells are activated to secrete perforin and granzyme, which induce cell death in B-cell leukemia. Blinatumomab is produced in the Chinese hamster ovary (CHO) cell expression system in consideration of post-translational modification (PTM) and immunogenicity. Although the characteristics of CHO cells are similar to those of other mammalian cells, it has low yield and high production cost. In this study, we developed the yeast species\n                    <jats:italic>Pichia pastoris<\/jats:italic>\n                    -produced BiTE antibody targeting CD19 and CD3\u03b5, herein called\n                    <jats:italic>P. pastoris<\/jats:italic>\n                    (p)-blinatumomab, which is in-house produced blinatumomab. Approximately 1.2\u00a0mg\/L CD19\/CD3\u03b5-targeting BiTE antibody (p-blinatumomab) was produced from the\n                    <jats:italic>P. pastoris<\/jats:italic>\n                    by transformation with the pHIL-S1 plasmid vector encoding the sequence of blinatumomab, which was 2.4 times higher than that produced in the CHO cell expression system based on the previous study. Enzyme-linked immunosorbent assay verified that the antigen binding affinity of the purified p-blinatumomab protein for CD19 and CD3\u03b5 antigens was 1.86 and 1.43 times higher than that of the commercial blinatumomab, BLINCYTO, respectively. In addition, flow cytometry confirmed that p-blinatumomab had a similar antigen binding affinity to CD19- and CD3\u03b5-overexpressing cells compared to the BLINCYTO. In conclusion, we suggest that\n                    <jats:italic>P. pastoris<\/jats:italic>\n                    is a useful system for expressing and producing BiTE antibody platforms for therapeutic drugs.\n                  <\/jats:p>","DOI":"10.1038\/s41598-025-21889-4","type":"journal-article","created":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T13:38:00Z","timestamp":1761831480000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Production system establishment and in vitro study of CD19\/CD3\u03b5 bite antibody secreted from Pichia pastoris"],"prefix":"10.1038","volume":"15","author":[{"given":"Boram","family":"Kim","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kyu Tae","family":"Byun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seung Hyeon","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"So Yeong","family":"Cheon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chan Gil","family":"Kim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,10,30]]},"reference":[{"key":"21889_CR1","doi-asserted-by":"publisher","first-page":"1146","DOI":"10.1016\/S0140-6736(19)33018-1","volume":"395","author":"F Malard","year":"2020","unstructured":"Malard, F. & Mohty, M. Acute lymphoblastic leukaemia. Lancet 395, 1146\u20131162. https:\/\/doi.org\/10.1016\/S0140-6736(19)33018-1 (2020).","journal-title":"Lancet"},{"key":"21889_CR2","doi-asserted-by":"publisher","first-page":"e577","DOI":"10.1038\/bcj.2017.53","volume":"7","author":"T Terwilliger","year":"2017","unstructured":"Terwilliger, T. & Abdul-Hay, M. Acute lymphoblastic leukemia: a comprehensive review and 2017 update. Blood Cancer J. 7, e577. https:\/\/doi.org\/10.1038\/bcj.2017.53 (2017).","journal-title":"Blood Cancer J."},{"doi-asserted-by":"publisher","unstructured":"Pagliaro, L. et al. Acute lymphoblastic leukaemia. Nat. Rev. Dis. Primers. 10 https:\/\/doi.org\/10.1038\/s41572-024-00525-x (2024).","key":"21889_CR3","DOI":"10.1038\/s41572-024-00525-x"},{"doi-asserted-by":"publisher","unstructured":"Bassan, R., Bourquin, J. P., DeAngelo, D. J. & Chiaretti, S. New approaches to the management of adult acute lymphoblastic leukemia. J. Clin. Oncol. JCO2017773648 https:\/\/doi.org\/10.1200\/JCO.2017.77.3648 (2018).","key":"21889_CR4","DOI":"10.1200\/JCO.2017.77.3648"},{"key":"21889_CR5","doi-asserted-by":"publisher","first-page":"12398","DOI":"10.1038\/s41598-021-91784-1","volume":"11","author":"YH Chen","year":"2021","unstructured":"Chen, Y. H., Wang, Y., Liao, C. H. & Hsu, S. C. The potential of adoptive transfer of gamma9delta2 T cells to enhance blinatumomab\u2019s antitumor activity against B-cell malignancy. Sci. Rep. 11, 12398. https:\/\/doi.org\/10.1038\/s41598-021-91784-1 (2021).","journal-title":"Sci. Rep."},{"key":"21889_CR6","doi-asserted-by":"publisher","first-page":"111356","DOI":"10.1016\/j.isci.2024.111356","volume":"27","author":"Y Hu","year":"2024","unstructured":"Hu, Y. et al. Global, regional, and National burden of acute lymphoblastic leukemia in children: epidemiological trends analysis from 1990 to 2021. iScience 27, 111356. https:\/\/doi.org\/10.1016\/j.isci.2024.111356 (2024).","journal-title":"iScience"},{"doi-asserted-by":"publisher","unstructured":"Tian, Z., Liu, M., Zhang, Y. & Wang, X. Bispecific T cell engagers: an emerging therapy for management of hematologic malignancies. J. Hematol. Oncol. 14 https:\/\/doi.org\/10.1186\/s13045-021-01084-4 (2021).","key":"21889_CR7","DOI":"10.1186\/s13045-021-01084-4"},{"key":"21889_CR8","doi-asserted-by":"publisher","first-page":"3192","DOI":"10.1002\/cncr.32909","volume":"126","author":"H Einsele","year":"2020","unstructured":"Einsele, H. et al. The bite (bispecific T-cell engager) platform: development and future potential of a targeted immuno-oncology therapy across tumor types. Cancer 126, 3192\u20133201. https:\/\/doi.org\/10.1002\/cncr.32909 (2020).","journal-title":"Cancer"},{"key":"21889_CR9","doi-asserted-by":"publisher","first-page":"5","DOI":"10.2147\/CPAA.S42689","volume":"5","author":"CA Portell","year":"2013","unstructured":"Portell, C. A., Wenzell, C. M. & Advani, A. S. Clinical and Pharmacologic aspects of blinatumomab in the treatment of B-cell acute lymphoblastic leukemia. Clin. Pharmacol. 5, 5\u201311. https:\/\/doi.org\/10.2147\/CPAA.S42689 (2013).","journal-title":"Clin. Pharmacol."},{"key":"21889_CR10","doi-asserted-by":"publisher","first-page":"895","DOI":"10.1517\/14712598.2015.1041912","volume":"15","author":"B Rogala","year":"2015","unstructured":"Rogala, B. et al. Blinatumomab: enlisting serial killer T-cells in the war against hematologic malignancies. Expert Opin. Biol. Ther. 15, 895\u2013908. https:\/\/doi.org\/10.1517\/14712598.2015.1041912 (2015).","journal-title":"Expert Opin. Biol. Ther."},{"key":"21889_CR11","doi-asserted-by":"publisher","first-page":"4035","DOI":"10.1158\/1078-0432.CCR-15-0612","volume":"21","author":"D Przepiorka","year":"2015","unstructured":"Przepiorka, D. et al. FDA approval: blinatumomab. Clin. Cancer Res. 21, 4035\u20134039. https:\/\/doi.org\/10.1158\/1078-0432.CCR-15-0612 (2015).","journal-title":"Clin. Cancer Res."},{"key":"21889_CR12","doi-asserted-by":"publisher","first-page":"107757","DOI":"10.1016\/j.biotechadv.2021.107757","volume":"49","author":"L Bryan","year":"2021","unstructured":"Bryan, L., Clynes, M. & Meleady, P. The emerging role of cellular post-translational modifications in modulating growth and productivity of Recombinant Chinese hamster ovary cells. Biotechnol. Adv. 49, 107757. https:\/\/doi.org\/10.1016\/j.biotechadv.2021.107757 (2021).","journal-title":"Biotechnol. Adv."},{"key":"21889_CR13","doi-asserted-by":"publisher","first-page":"1337","DOI":"10.1111\/jcpt.13741","volume":"47","author":"P Mocquot","year":"2022","unstructured":"Mocquot, P., Mossazadeh, Y., Lapierre, L., Pineau, F. & Despas, F. The Pharmacology of blinatumomab: state of the Art on pharmacodynamics, pharmacokinetics, adverse drug reactions and evaluation in clinical trials. J. Clin. Pharm. Ther. 47, 1337\u20131351. https:\/\/doi.org\/10.1111\/jcpt.13741 (2022).","journal-title":"J. Clin. Pharm. Ther."},{"key":"21889_CR14","doi-asserted-by":"publisher","first-page":"182","DOI":"10.1007\/s00253-024-13008-6","volume":"108","author":"SL Geng","year":"2024","unstructured":"Geng, S. L. et al. Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells. Appl. Microbiol. Biotechnol. 108, 182. https:\/\/doi.org\/10.1007\/s00253-024-13008-6 (2024).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"21889_CR15","doi-asserted-by":"publisher","first-page":"880155","DOI":"10.3389\/fbioe.2022.880155","volume":"10","author":"ZM Li","year":"2022","unstructured":"Li, Z. M., Fan, Z. L., Wang, X. Y. & Wang, T. Y. Factors affecting the expression of Recombinant protein and improvement strategies in Chinese hamster ovary cells. Front. Bioeng. Biotechnol. 10, 880155. https:\/\/doi.org\/10.3389\/fbioe.2022.880155 (2022).","journal-title":"Front. Bioeng. Biotechnol."},{"key":"21889_CR16","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1007\/978-3-540-70868-1_4","volume":"332","author":"K Ko","year":"2009","unstructured":"Ko, K., Brodzik, R. & Steplewski, Z. Production of antibodies in plants: approaches and perspectives. Curr. Top. Microbiol. Immunol. 332, 55\u201378. https:\/\/doi.org\/10.1007\/978-3-540-70868-1_4 (2009).","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"21889_CR17","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1016\/j.nbt.2018.04.006","volume":"44","author":"K Lebozec","year":"2018","unstructured":"Lebozec, K., Jandrot-Perrus, M., Avenard, G., Favre-Bulle, O. & Billiald, P. Quality and cost assessment of a Recombinant antibody fragment produced from mammalian, yeast and prokaryotic host cells: A case study prior to pharmaceutical development. N Biotechnol. 44, 31\u201340. https:\/\/doi.org\/10.1016\/j.nbt.2018.04.006 (2018).","journal-title":"N Biotechnol."},{"key":"21889_CR18","doi-asserted-by":"publisher","first-page":"526","DOI":"10.1002\/biot.201300305","volume":"9","author":"A Maccani","year":"2014","unstructured":"Maccani, A. et al. Pichia pastoris secretes Recombinant proteins less efficiently than Chinese hamster ovary cells but allows higher space-time yields for less complex proteins. Biotechnol. J. 9, 526\u2013537. https:\/\/doi.org\/10.1002\/biot.201300305 (2014).","journal-title":"Biotechnol. J."},{"key":"21889_CR19","doi-asserted-by":"publisher","first-page":"5867","DOI":"10.1002\/jcp.29583","volume":"235","author":"M Karbalaei","year":"2020","unstructured":"Karbalaei, M., Rezaee, S. A. & Farsiani, H. Pichia pastoris: A highly successful expression system for optimal synthesis of heterologous proteins. J. Cell. Physiol. 235, 5867\u20135881. https:\/\/doi.org\/10.1002\/jcp.29583 (2020).","journal-title":"J. Cell. Physiol."},{"key":"21889_CR20","doi-asserted-by":"publisher","first-page":"587","DOI":"10.4014\/jmb.1210.10063","volume":"23","author":"AM Goncalves","year":"2013","unstructured":"Goncalves, A. M. et al. Pichia pastoris: a Recombinant microfactory for antibodies and human membrane proteins. J. Microbiol. Biotechnol. 23, 587\u2013601. https:\/\/doi.org\/10.4014\/jmb.1210.10063 (2013).","journal-title":"J. Microbiol. Biotechnol."},{"key":"21889_CR21","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1007\/978-1-4939-7312-5_3","volume":"1674","author":"A Nylen","year":"2018","unstructured":"Nylen, A. & Chen, M. T. Production of Full-Length antibody by Pichia pastoris. Methods Mol. Biol. 1674, 37\u201348. https:\/\/doi.org\/10.1007\/978-1-4939-7312-5_3 (2018).","journal-title":"Methods Mol. Biol."},{"key":"21889_CR22","doi-asserted-by":"publisher","first-page":"11225","DOI":"10.1038\/s41598-022-15086-w","volume":"12","author":"D Wollborn","year":"2022","unstructured":"Wollborn, D. et al. Predicting high Recombinant protein producer strains of Pichia pastoris Mut(S) using the oxygen transfer rate as an indicator of metabolic burden. Sci. Rep. 12, 11225. https:\/\/doi.org\/10.1038\/s41598-022-15086-w (2022).","journal-title":"Sci. Rep."},{"key":"21889_CR23","doi-asserted-by":"publisher","first-page":"183111","DOI":"10.1016\/j.bbamem.2019.183111","volume":"1862","author":"D Dutta","year":"2020","unstructured":"Dutta, D., Esmaili, M., Overduin, M. & Fliegel, L. Expression and detergent free purification and reconstitution of the plant plasma membrane Na(+)\/H(+) antiporter SOS1 overexpressed in Pichia pastoris. Biochim. Biophys. Acta Biomembr. 1862, 183111. https:\/\/doi.org\/10.1016\/j.bbamem.2019.183111 (2020).","journal-title":"Biochim. Biophys. Acta Biomembr."},{"key":"21889_CR24","doi-asserted-by":"publisher","first-page":"711","DOI":"10.1007\/s10529-013-1411-3","volume":"36","author":"Y Chen","year":"2014","unstructured":"Chen, Y., Li, Y., Liu, P., Sun, Q. & Liu, Z. Optimized expression in Pichia pastoris eliminates common protein contaminants from subsequent His-tag purification. Biotechnol. Lett. 36, 711\u2013718. https:\/\/doi.org\/10.1007\/s10529-013-1411-3 (2014).","journal-title":"Biotechnol. Lett."},{"key":"21889_CR25","doi-asserted-by":"publisher","first-page":"55","DOI":"10.22088\/IJMCM.BUMS.8.1.55","volume":"8","author":"F Naddafi","year":"2019","unstructured":"Naddafi, F. et al. The epigenetic regulation of blinatumomab gene expression: tumor Cell-dependent T cell response against lymphoma cells and cytotoxic activity. Int. J. Mol. Cell. Med. 8, 55\u201366. https:\/\/doi.org\/10.22088\/IJMCM.BUMS.8.1.55 (2019).","journal-title":"Int. J. Mol. Cell. Med."},{"key":"21889_CR26","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1186\/1475-2859-11-91","volume":"11","author":"MT Chen","year":"2012","unstructured":"Chen, M. T. et al. Generation of diploid Pichia pastoris strains by mating and their application for Recombinant protein production. Microb. Cell. Fact. 11, 91. https:\/\/doi.org\/10.1186\/1475-2859-11-91 (2012).","journal-title":"Microb. Cell. Fact."},{"key":"21889_CR27","doi-asserted-by":"publisher","first-page":"124","DOI":"10.1186\/s12934-021-01617-z","volume":"20","author":"A Madhavan","year":"2021","unstructured":"Madhavan, A. et al. Customized yeast cell factories for biopharmaceuticals: from cell engineering to process scale up. Microb. Cell. Fact. 20, 124. https:\/\/doi.org\/10.1186\/s12934-021-01617-z (2021).","journal-title":"Microb. Cell. Fact."},{"key":"21889_CR28","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1016\/j.jim.2012.08.015","volume":"386","author":"MC Cukan","year":"2012","unstructured":"Cukan, M. C. et al. Binding of DC-SIGN to glycoproteins expressed in glycoengineered Pichia pastoris. J. Immunol. Methods. 386, 34\u201342. https:\/\/doi.org\/10.1016\/j.jim.2012.08.015 (2012).","journal-title":"J. Immunol. Methods"},{"key":"21889_CR29","doi-asserted-by":"publisher","first-page":"752","DOI":"10.1016\/j.bbrc.2018.06.071","volume":"503","author":"H Nakamura","year":"2018","unstructured":"Nakamura, H., Oda-Ueda, N., Ueda, T. & Ohkuri, T. Introduction of a glycosylation site in the constant region decreases the aggregation of adalimumab fab. Biochem. Biophys. Res. Commun. 503, 752\u2013756. https:\/\/doi.org\/10.1016\/j.bbrc.2018.06.071 (2018).","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"21889_CR30","doi-asserted-by":"publisher","first-page":"123407","DOI":"10.1016\/j.ijbiomac.2023.123407","volume":"233","author":"A Beygmoradi","year":"2023","unstructured":"Beygmoradi, A., Homaei, A., Hemmati, R. & Fernandes, P. Recombinant protein expression: challenges in production and folding related matters. Int. J. Biol. Macromol. 233, 123407. https:\/\/doi.org\/10.1016\/j.ijbiomac.2023.123407 (2023).","journal-title":"Int. J. Biol. Macromol."},{"key":"21889_CR31","doi-asserted-by":"publisher","first-page":"6245","DOI":"10.1074\/jbc.M115.692053","volume":"291","author":"X Wang","year":"2016","unstructured":"Wang, X. et al. Mit1 transcription factor mediates methanol signaling and regulates the alcohol oxidase 1 (AOX1) promoter in Pichia pastoris. J. Biol. Chem. 291, 6245\u20136261. https:\/\/doi.org\/10.1074\/jbc.M115.692053 (2016).","journal-title":"J. Biol. Chem."},{"key":"21889_CR32","doi-asserted-by":"publisher","first-page":"41850","DOI":"10.1038\/srep41850","volume":"7","author":"J Wang","year":"2017","unstructured":"Wang, J. et al. Methanol-Independent protein expression by AOX1 promoter with trans-Acting elements engineering and Glucose-Glycerol-Shift induction in Pichia pastoris. Sci. Rep. 7, 41850. https:\/\/doi.org\/10.1038\/srep41850 (2017).","journal-title":"Sci. Rep."},{"key":"21889_CR33","doi-asserted-by":"publisher","first-page":"18439","DOI":"10.1038\/srep18439","volume":"6","author":"WC Liu","year":"2016","unstructured":"Liu, W. C. et al. Scaling-up fermentation of Pichia pastoris to demonstration-scale using new methanol-feeding strategy and increased air pressure instead of pure oxygen supplement. Sci. Rep. 6, 18439. https:\/\/doi.org\/10.1038\/srep18439 (2016).","journal-title":"Sci. Rep."},{"key":"21889_CR34","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1016\/j.copbio.2017.12.010","volume":"53","author":"KR Love","year":"2018","unstructured":"Love, K. R., Dalvie, N. C. & Love, J. C. The yeast stands alone: the future of protein biologic production. Curr. Opin. Biotechnol. 53, 50\u201358. https:\/\/doi.org\/10.1016\/j.copbio.2017.12.010 (2018).","journal-title":"Curr. Opin. Biotechnol."},{"key":"21889_CR35","doi-asserted-by":"publisher","first-page":"5301","DOI":"10.1007\/s00253-014-5732-5","volume":"98","author":"M Ahmad","year":"2014","unstructured":"Ahmad, M., Hirz, M., Pichler, H. & Schwab, H. Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production. Appl. Microbiol. Biotechnol. 98, 5301\u20135317. https:\/\/doi.org\/10.1007\/s00253-014-5732-5 (2014).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"21889_CR36","doi-asserted-by":"publisher","first-page":"363","DOI":"10.1007\/s00253-004-1656-9","volume":"65","author":"FR Schmidt","year":"2004","unstructured":"Schmidt, F. R. Recombinant expression systems in the pharmaceutical industry. Appl. Microbiol. Biotechnol. 65, 363\u2013372. https:\/\/doi.org\/10.1007\/s00253-004-1656-9 (2004).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"21889_CR37","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1586\/eci.09.60","volume":"6","author":"JA Bernstein","year":"2010","unstructured":"Bernstein, J. A. & Qazi, M. Ecallantide: its pharmacology, pharmacokinetics, clinical efficacy and tolerability. Expert Rev. Clin. Immunol. 6, 29\u201339. https:\/\/doi.org\/10.1586\/eci.09.60 (2010).","journal-title":"Expert Rev. Clin. Immunol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-21889-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-21889-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-21889-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T13:38:01Z","timestamp":1761831481000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-21889-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,30]]},"references-count":37,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["21889"],"URL":"https:\/\/doi.org\/10.1038\/s41598-025-21889-4","relation":{},"ISSN":["2045-2322"],"issn-type":[{"type":"electronic","value":"2045-2322"}],"subject":[],"published":{"date-parts":[[2025,10,30]]},"assertion":[{"value":"26 May 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 September 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"30 October 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"38061"}}