{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,6]],"date-time":"2026-01-06T13:21:20Z","timestamp":1767705680000,"version":"build-2065373602"},"reference-count":49,"publisher":"Elsevier BV","license":[{"start":{"date-parts":[[2023,6,1]],"date-time":"2023-06-01T00:00:00Z","timestamp":1685577600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/tdm\/userlicense\/1.0\/"},{"start":{"date-parts":[[2023,6,1]],"date-time":"2023-06-01T00:00:00Z","timestamp":1685577600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/legal\/tdmrep-license"},{"start":{"date-parts":[[2023,3,28]],"date-time":"2023-03-28T00:00:00Z","timestamp":1679961600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["elsevier.com","sciencedirect.com"],"crossmark-restriction":true},"short-container-title":["European Journal of Pharmaceutics and Biopharmaceutics"],"published-print":{"date-parts":[[2023,6]]},"DOI":"10.1016\/j.ejpb.2023.03.012","type":"journal-article","created":{"date-parts":[[2023,4,1]],"date-time":"2023-04-01T11:18:17Z","timestamp":1680347897000},"page":"1-11","update-policy":"https:\/\/doi.org\/10.1016\/elsevier_cm_policy","source":"Crossref","is-referenced-by-count":3,"special_numbering":"C","title":["Isothermal denaturation fluorimetry vs differential scanning fluorimetry as tools for screening of stabilizers for protein freeze-drying: Human phenylalanine hydroxylase as the case study"],"prefix":"10.1016","volume":"187","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2946-9342","authenticated-orcid":false,"given":"Paula","family":"Leandro","sequence":"first","affiliation":[]},{"given":"Paulo R.","family":"Lino","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4103-3752","authenticated-orcid":false,"given":"Raquel","family":"Lopes","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0163-3235","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Leandro","sequence":"additional","affiliation":[]},{"given":"Mariana P.","family":"Amaro","sequence":"additional","affiliation":[]},{"given":"Paulo","family":"Sousa","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5244-8059","authenticated-orcid":false,"given":"Jo\u00e3o B.","family":"Vicente","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7807-4726","authenticated-orcid":false,"given":"Ant\u00f3nio J.","family":"Almeida","sequence":"additional","affiliation":[]}],"member":"78","reference":[{"key":"10.1016\/j.ejpb.2023.03.012_b0005","doi-asserted-by":"crossref","first-page":"649","DOI":"10.1042\/EBC20190042","article-title":"Uncovering protein structure","volume":"64","author":"Stollar","year":"2020","journal-title":"Essays Biochem."},{"key":"10.1016\/j.ejpb.2023.03.012_b0010","series-title":"Pharmaceutical Biotechnology: Fundamentals and Applications","first-page":"83","article-title":"Formulation of Biologics Including Biopharmaceutical Considerations","author":"Crommelin","year":"2019"},{"key":"10.1016\/j.ejpb.2023.03.012_b0015","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.addr.2014.10.005","article-title":"Structural and functional stabilization of protein entities: state-of-the-art","volume":"93","author":"Balcao","year":"2015","journal-title":"Adv. Drug Deliv. Rev."},{"key":"10.1016\/j.ejpb.2023.03.012_b0020","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.addr.2015.05.006","article-title":"Stabilization of proteins in solid form","volume":"93","author":"Cicerone","year":"2015","journal-title":"Adv. Drug Deliv. Rev."},{"key":"10.1016\/j.ejpb.2023.03.012_b0025","doi-asserted-by":"crossref","DOI":"10.3390\/pharmaceutics10030131","article-title":"Drying Technologies for the Stability and Bioavailability of Biopharmaceuticals","volume":"10","author":"Emami","year":"2018","journal-title":"Pharmaceutics"},{"key":"10.1016\/j.ejpb.2023.03.012_b0030","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/j.ijbiomac.2020.11.188","article-title":"Instability of therapeutic proteins - An overview of stresses, stabilization mechanisms and analytical techniques involved in lyophilized proteins","volume":"167","author":"Butreddy","year":"2021","journal-title":"Int. J. Biol. Macromol."},{"key":"10.1016\/j.ejpb.2023.03.012_b0035","doi-asserted-by":"crossref","DOI":"10.1016\/j.ijpharm.2020.119029","article-title":"Excipients in freeze-dried biopharmaceuticals: Contributions toward formulation stability and lyophilisation cycle optimisation","volume":"576","author":"Bjelosevic","year":"2020","journal-title":"Int. J. Pharm."},{"key":"10.1016\/j.ejpb.2023.03.012_b0040","doi-asserted-by":"crossref","first-page":"3373","DOI":"10.1080\/07373937.2022.2048847","article-title":"A comprehensive review on stability of therapeutic proteins treated by freeze-drying: induced stresses and stabilization mechanisms involved in processing","volume":"40","author":"Wang","year":"2022","journal-title":"Drying Technology"},{"key":"10.1016\/j.ejpb.2023.03.012_b0045","doi-asserted-by":"crossref","first-page":"763","DOI":"10.1016\/j.bbapap.2013.01.020","article-title":"Unraveling protein stabilization mechanisms: vitrification and water replacement in a glass transition temperature controlled system","volume":"1834","author":"Grasmeijer","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"10.1016\/j.ejpb.2023.03.012_b0050","doi-asserted-by":"crossref","first-page":"2116","DOI":"10.1016\/j.xphs.2020.03.022","article-title":"The Ice-Water Interface and Protein Stability: A Review","volume":"109","author":"Arsiccio","year":"2020","journal-title":"J. Pharm. Sci."},{"key":"10.1016\/j.ejpb.2023.03.012_b0055","doi-asserted-by":"crossref","first-page":"1883","DOI":"10.1007\/s11095-007-9312-6","article-title":"Stabilizing effect of four types of disaccharide on the enzymatic activity of freeze-dried lactate dehydrogenase: step by step evaluation from freezing to storage","volume":"24","author":"Kawai","year":"2007","journal-title":"Pharm. Res."},{"key":"10.1016\/j.ejpb.2023.03.012_b0060","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.1517\/17425240903199143","article-title":"Recent trends in stabilising peptides and proteins in pharmaceutical formulation - considerations in the choice of excipients","volume":"6","author":"Jorgensen","year":"2009","journal-title":"Expert Opin. Drug Deliv."},{"key":"10.1016\/j.ejpb.2023.03.012_b0065","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S0079-6603(08)00603-X","article-title":"Rescuing proteins of low kinetic stability by chaperones and natural ligands phenylketonuria, a case study","volume":"83","author":"Martinez","year":"2008","journal-title":"Prog. Mol. Biol. Transl. Sci."},{"key":"10.1016\/j.ejpb.2023.03.012_b0070","doi-asserted-by":"crossref","DOI":"10.3390\/nano12060893","article-title":"Impact of Fluorinated Ionic Liquids on Human Phenylalanine Hydroxylase-A Potential Drug Delivery System","volume":"12","author":"Alves","year":"2022","journal-title":"Nanomaterials (Basel)"},{"key":"10.1016\/j.ejpb.2023.03.012_b0075","doi-asserted-by":"crossref","DOI":"10.3390\/ijms22157987","article-title":"Systematic Modification and Evaluation of Enzyme-Loaded Chitosan Nanoparticles","volume":"22","author":"Lino","year":"2021","journal-title":"Int. J. Mol. Sci."},{"key":"10.1016\/j.ejpb.2023.03.012_b0080","doi-asserted-by":"crossref","DOI":"10.3390\/pharmaceutics13030329","article-title":"In Silico and In Vitro Tailoring of a Chitosan Nanoformulation of a Human Metabolic Enzyme","volume":"13","author":"Lino","year":"2021","journal-title":"Pharmaceutics"},{"key":"10.1016\/j.ejpb.2023.03.012_b0085","doi-asserted-by":"crossref","first-page":"2137","DOI":"10.1021\/cr980450y","article-title":"Structural Insight into the Aromatic Amino Acid Hydroxylases and Their Disease-Related Mutant Forms","volume":"99","author":"Flatmark","year":"1999","journal-title":"Chem. Rev."},{"key":"10.1016\/j.ejpb.2023.03.012_b0090","doi-asserted-by":"crossref","first-page":"997","DOI":"10.1046\/j.1432-1327.2001.01958.x","volume":"268","author":"Bjorgo","year":"2001","journal-title":"Eur. J. Biochem."},{"key":"10.1016\/j.ejpb.2023.03.012_b0095","doi-asserted-by":"crossref","first-page":"23748","DOI":"10.1038\/srep23748","article-title":"Structural basis for ligand-dependent dimerization of phenylalanine hydroxylase regulatory domain","volume":"6","author":"Patel","year":"2016","journal-title":"Sci. Rep."},{"key":"10.1016\/j.ejpb.2023.03.012_b0100","doi-asserted-by":"crossref","first-page":"2394","DOI":"10.1073\/pnas.1516967113","article-title":"First structure of full-length mammalian phenylalanine hydroxylase reveals the architecture of an autoinhibited tetramer","volume":"113","author":"Arturo","year":"2016","journal-title":"Proc. Natl. Acad. Sci. U S A"},{"key":"10.1016\/j.ejpb.2023.03.012_b0105","doi-asserted-by":"crossref","first-page":"13615","DOI":"10.1038\/s41598-019-49944-x","article-title":"Structure of full-length wild-type human phenylalanine hydroxylase by small angle X-ray scattering reveals substrate-induced conformational stability","volume":"9","author":"Tom\u00e9","year":"2019","journal-title":"Sci. Rep."},{"key":"10.1016\/j.ejpb.2023.03.012_b0110","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1006\/mgme.2000.2970","article-title":"The V388M mutation results in a kinetic variant form of phenylalanine hydroxylase","volume":"69","author":"Leandro","year":"2000","journal-title":"Mol. Genet. Metab."},{"key":"10.1016\/j.ejpb.2023.03.012_b0115","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1007\/s12010-009-8862-y","article-title":"Polyol additives modulate the in vitro stability and activity of recombinant human phenylalanine hydroxylase","volume":"162","author":"Nascimento","year":"2010","journal-title":"Appl. Biochem. Biotechnol."},{"key":"10.1016\/j.ejpb.2023.03.012_b0120","doi-asserted-by":"crossref","first-page":"61022","DOI":"10.1039\/C4RA10306H","article-title":"Phenylalanine iminoboronates as new phenylalanine hydroxylase modulators","volume":"4","author":"Montalbano","year":"2014","journal-title":"RSC Advances"},{"key":"10.1016\/j.ejpb.2023.03.012_b0125","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1177\/1087057108317825","article-title":"Application of high-throughput isothermal denaturation to assess protein stability and screen for ligands","volume":"13","author":"Senisterra","year":"2008","journal-title":"J. Biomol. Screen"},{"key":"10.1016\/j.ejpb.2023.03.012_b0130","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem."},{"key":"10.1016\/j.ejpb.2023.03.012_b0135","doi-asserted-by":"crossref","DOI":"10.3390\/biom11030462","article-title":"Modulation of Human Phenylalanine Hydroxylase by 3-Hydroxyquinolin-2(1H)-One Derivatives","volume":"11","author":"Lopes","year":"2021","journal-title":"Biomolecules"},{"key":"10.1016\/j.ejpb.2023.03.012_b0140","doi-asserted-by":"crossref","first-page":"3926","DOI":"10.1016\/j.jchromb.2009.09.045","article-title":"Determination of phenylalanine and tyrosine in plasma and dried blood samples using HPLC with fluorescence detection","volume":"877","author":"Kand'ar","year":"2009","journal-title":"J. Chromatogr. B Analyt. Technol. Biomed. Life Sci."},{"key":"10.1016\/j.ejpb.2023.03.012_b0145","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1007\/s00249-009-0403-7","article-title":"Study of kinetics of thermal aggregation of mitochondrial aspartate aminotransferase by dynamic light scattering: protective effect of alpha-crystallin","volume":"38","author":"Golub","year":"2009","journal-title":"Eur. Biophys. J."},{"key":"10.1016\/j.ejpb.2023.03.012_b0150","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1039\/B814377C","article-title":"High throughput methods of assessing protein stability and aggregation","volume":"5","author":"Senisterra","year":"2009","journal-title":"Mol. Biosyst."},{"key":"10.1016\/j.ejpb.2023.03.012_b0155","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/j.ajhg.2008.05.013","article-title":"Loss of function in phenylketonuria is caused by impaired molecular motions and conformational instability","volume":"83","author":"Gersting","year":"2008","journal-title":"Am. J. Hum. Genet."},{"key":"10.1016\/j.ejpb.2023.03.012_b0160","doi-asserted-by":"crossref","first-page":"7573","DOI":"10.1021\/bi0160720","article-title":"L-phenylalanine binding and domain organization in human phenylalanine hydroxylase: a differential scanning calorimetry study","volume":"41","author":"Thorolfsson","year":"2002","journal-title":"Biochemistry"},{"key":"10.1016\/j.ejpb.2023.03.012_b0165","doi-asserted-by":"crossref","first-page":"1136","DOI":"10.1038\/nbt.4305","article-title":"Biopharmaceutical benchmarks 2018","volume":"36","author":"Walsh","year":"2018","journal-title":"Nat. Biotechnol."},{"key":"10.1016\/j.ejpb.2023.03.012_b0170","doi-asserted-by":"crossref","first-page":"e0181748","DOI":"10.1371\/journal.pone.0181748","article-title":"THPdb: Database of FDA-approved peptide and protein therapeutics","volume":"12","author":"Usmani","year":"2017","journal-title":"PLoS One"},{"key":"10.1016\/j.ejpb.2023.03.012_b0175","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.ejpb.2018.07.011","article-title":"Parenteral protein formulations: An overview of approved products within the European Union","volume":"131","author":"Gervasi","year":"2018","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"10.1016\/j.ejpb.2023.03.012_b0180","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1517\/17460441.2013.806479","article-title":"Recent developments in the use of differential scanning fluorometry in protein and small molecule discovery and characterization","volume":"8","author":"Simeonov","year":"2013","journal-title":"Expert Opin Drug Discov."},{"key":"10.1016\/j.ejpb.2023.03.012_b0185","doi-asserted-by":"crossref","first-page":"15835","DOI":"10.1073\/pnas.0605224103","article-title":"Chemical screening methods to identify ligands that promote protein stability, protein crystallization, and structure determination","volume":"103","author":"Vedadi","year":"2006","journal-title":"Proc. Natl. Acad. Sci. U S A"},{"key":"10.1016\/j.ejpb.2023.03.012_b0190","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.jpba.2013.01.006","article-title":"Differential scanning fluorimetry: rapid screening of formulations that promote the stability of reference preparations","volume":"77","author":"Malik","year":"2013","journal-title":"J. Pharm. Biomed. Anal."},{"key":"10.1016\/j.ejpb.2023.03.012_b0195","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.ejpb.2018.01.004","article-title":"Isothermal chemical denaturation as a complementary tool to overcome limitations of thermal differential scanning fluorimetry in predicting physical stability of protein formulations","volume":"125","author":"Svilenov","year":"2018","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"10.1016\/j.ejpb.2023.03.012_b0200","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.ejpb.2006.09.009","article-title":"High throughput screening of protein formulation stability: practical considerations","volume":"65","author":"Capelle","year":"2007","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"10.1016\/j.ejpb.2023.03.012_b0205","doi-asserted-by":"crossref","first-page":"33251","DOI":"10.1074\/jbc.274.47.33251","article-title":"Urea-induced denaturation of human phenylalanine hydroxylase","volume":"274","author":"Kleppe","year":"1999","journal-title":"J. Biol. Chem."},{"key":"10.1016\/j.ejpb.2023.03.012_b0210","doi-asserted-by":"crossref","DOI":"10.1016\/j.ijpharm.2022.121492","article-title":"Formulation of spray dried enzymes for dry powder inhalers: An integrated methodology","volume":"615","author":"Fernandes","year":"2022","journal-title":"Int. J. Pharm."},{"key":"10.1016\/j.ejpb.2023.03.012_b0215","doi-asserted-by":"crossref","first-page":"2283","DOI":"10.1002\/jps.10154","article-title":"Excipient-drug interactions in parenteral formulations","volume":"91","author":"Akers","year":"2002","journal-title":"J. Pharm. Sci."},{"key":"10.1016\/j.ejpb.2023.03.012_b0220","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1248\/cpb.42.5","article-title":"Effect of mannitol crystallinity on the stabilization of enzymes during freeze-drying","volume":"42","author":"Izutsu","year":"1994","journal-title":"Chem. Pharm. Bull. (Tokyo)"},{"key":"10.1016\/j.ejpb.2023.03.012_b0225","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1006\/abbi.1998.0948","article-title":"Effects of additives on the stability of recombinant human factor XIII during freeze-drying and storage in the dried solid","volume":"360","author":"Kreilgaard","year":"1998","journal-title":"Arch. Biochem. Biophys."},{"key":"10.1016\/j.ejpb.2023.03.012_b0230","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.ijpharm.2015.03.051","article-title":"Controlling protein stability: Mechanisms revealed using formulations of arginine, glycine and guanidinium HCl with three globular proteins","volume":"486","author":"Platts","year":"2015","journal-title":"Int. J. Pharm."},{"key":"10.1016\/j.ejpb.2023.03.012_b0235","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1007\/s00726-013-1528-7","article-title":"L-arginine induces protein aggregation and transformation of supramolecular structures of the aggregates","volume":"45","author":"Smirnova","year":"2013","journal-title":"Amino Acids"},{"key":"10.1016\/j.ejpb.2023.03.012_b0240","doi-asserted-by":"crossref","first-page":"4444","DOI":"10.1002\/jps.23332","article-title":"Local dynamics and their alteration by excipients modulate the global conformational stability of an lgG1 monoclonal antibody","volume":"101","author":"Thakkar","year":"2012","journal-title":"J. Pharm. Sci."},{"key":"10.1016\/j.ejpb.2023.03.012_b0245","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ijpharm.2012.04.040","article-title":"Immunogenicity of protein aggregates\u2013concerns and realities","volume":"431","author":"Wang","year":"2012","journal-title":"Int. J. Pharm."}],"container-title":["European Journal of Pharmaceutics and Biopharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0939641123000784?httpAccept=text\/xml","content-type":"text\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0939641123000784?httpAccept=text\/plain","content-type":"text\/plain","content-version":"vor","intended-application":"text-mining"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T16:50:50Z","timestamp":1760547050000},"score":1,"resource":{"primary":{"URL":"https:\/\/linkinghub.elsevier.com\/retrieve\/pii\/S0939641123000784"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6]]},"references-count":49,"alternative-id":["S0939641123000784"],"URL":"https:\/\/doi.org\/10.1016\/j.ejpb.2023.03.012","relation":{},"ISSN":["0939-6411"],"issn-type":[{"type":"print","value":"0939-6411"}],"subject":[],"published":{"date-parts":[[2023,6]]},"assertion":[{"value":"Elsevier","name":"publisher","label":"This article is maintained by"},{"value":"Isothermal denaturation fluorimetry vs differential scanning fluorimetry as tools for screening of stabilizers for protein freeze-drying: Human phenylalanine hydroxylase as the case study","name":"articletitle","label":"Article Title"},{"value":"European Journal of Pharmaceutics and Biopharmaceutics","name":"journaltitle","label":"Journal Title"},{"value":"https:\/\/doi.org\/10.1016\/j.ejpb.2023.03.012","name":"articlelink","label":"CrossRef DOI link to publisher maintained version"},{"value":"article","name":"content_type","label":"Content Type"},{"value":"\u00a9 2023 The Author(s). Published by Elsevier B.V.","name":"copyright","label":"Copyright"}]}}