{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T02:12:29Z","timestamp":1793067149500,"version":"4.5.0"},"reference-count":41,"publisher":"American Chemical Society (ACS)","issue":"4","license":[{"start":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T00:00:00Z","timestamp":1778544000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004543","name":"China Scholarship Council","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004543","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,19]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Integrating therapeutic proteins into lipid-based nanocarriers remains challenging due to fundamental incompatibilities between hydrophilic protein surfaces and lipophilic carrier matrices. Here, we demonstrate that controlled hydrophobic ion pairing (HIP) using lysozyme-sodium dodecyl sulfate (SDS) at stoichiometric ratios simultaneously improves thermal stability during pharmaceutical processing and lipid compatibility for nanocarrier integration. Building on our previous molecular characterization of lysozyme-SDS complexes, we investigated their application in nanostructured lipid carrier (NLC) formulations for oral protein delivery. SDS complexation preserved lysozyme enzymatic activity at processing-relevant temperatures and increased apparent lipid miscibility by 50% in both solid and liquid pharmaceutical lipid excipients. This enhanced compatibility translated directly into superior NLC performance, with complexed lysozyme achieving a 4-fold higher encapsulation efficiency (79.6 \u00b1 1.8% vs 18.4 \u00b1 4.3% for native protein) with improved batch-to-batch reproducibility. In simulated gastrointestinal conditions, lysozyme-SDS NLCs demonstrated sustained intestinal release (87.2 \u00b1 16.5% over 6 h) with 67.5 \u00b1 7.5% enzymatic activity retained compared to only 15.2 \u00b1 3.0% activity for native lysozyme formulations. Mechanistic analysis from differential scanning calorimetry and crystallinity measurements showed that SDS complexation induces a molten globule-like protein state that reduces lipid packing (56.8% vs 68.6% crystallinity for native lysozyme), enabling superior matrix integration while maintaining biological function. These findings establish that controlled surface modification through HIP provides a systematic approach to overcome protein\u2013lipid incompatibilities, offering a generalizable framework for developing lipid-based protein therapeutics.<\/jats:p>","DOI":"10.1021\/acsnanoscienceau.6c00033","type":"journal-article","created":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T10:23:39Z","timestamp":1778581419000},"page":"644-653","source":"Crossref","is-referenced-by-count":1,"title":["Hydrophobic\nIon Pairing Improves Protein Thermal Stability\nand Lipid Nanocarrier Integration through Controlled Surface Modification"],"prefix":"10.1021","volume":"6","author":[{"given":"Jiaming","family":"Mu","sequence":"first","affiliation":[{"name":"Queen\u2019s University Belfast , , ,","place":["Belfast, United Kingdom, BT9 7BL"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Sedough-Abbasian","sequence":"additional","affiliation":[{"name":"Queen\u2019s University Belfast , , ,","place":["Belfast, United Kingdom, BT9 7BL"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9106-9836","authenticated-orcid":true,"given":"Leran","family":"Mao","sequence":"additional","affiliation":[{"name":"Carnegie Mellon University , , , ,","place":["Pittsburgh, Pennsylvania, United States, 15213"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3436-4745","authenticated-orcid":true,"given":"Sheiliza","family":"Carmali","sequence":"additional","affiliation":[{"name":"Queen\u2019s University Belfast , , ,","place":["Belfast, United Kingdom, BT9 7BL"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,5,12]]},"reference":[{"issue":"1","key":"2026081909070729000_cit1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/S0005-2736(03)00056-7","article-title":"Lipid\u2013Protein\nInteractions in Biological Membranes: A Structural Perspective","volume":"1612","author":"Lee","year":"2003","journal-title":"Biochim. Biophys. Acta , Biomembr."},{"issue":"9","key":"2026081909070729000_cit2","doi-asserted-by":"publisher","first-page":"5775","DOI":"10.1021\/acs.chemrev.8b00451","article-title":"Emerging\nDiversity\nin Lipid\u2013Protein Interactions","volume":"119","author":"Corradi","year":"2019","journal-title":"Chem.\nRev."},{"issue":"1","key":"2026081909070729000_cit3","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1146\/annurev-biophys-090721-072718","article-title":"Lipid\u2013Protein\nInteractions\nin Plasma Membrane Organization and Function","volume":"51","author":"Sych","year":"2022","journal-title":"Annu. Rev. Biophys."},{"issue":"9","key":"2026081909070729000_cit4","doi-asserted-by":"publisher","first-page":"6086","DOI":"10.1021\/acs.chemrev.8b00608","article-title":"Characterization\nof Lipid\u2013Protein Interactions and Lipid-Mediated Modulation\nof Membrane Protein Function through Molecular Simulation","volume":"119","author":"Muller","year":"2019","journal-title":"Chem. Rev."},{"issue":"1","key":"2026081909070729000_cit5","doi-asserted-by":"publisher","first-page":"2411","DOI":"10.1038\/s41467-023-38039-x","article-title":"Engineering Protein-Based\nTherapeutics through Structural\nand Chemical Design","volume":"14","author":"Ebrahimi","year":"2023","journal-title":"Nat. Commun."},{"issue":"3","key":"2026081909070729000_cit6","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1038\/s41570-019-0079-1","article-title":"Contemporary\nApproaches to Site-Selective Protein Modification","volume":"3","author":"Hoyt","year":"2019","journal-title":"Nat. Rev. Chem."},{"issue":"2","key":"2026081909070729000_cit7","doi-asserted-by":"publisher","first-page":"188","DOI":"10.1023\/A:1011998014474","article-title":"Hydrophobic Ion\nPairing: Altering the Solubility Properties\nof Biomolecules","volume":"15","author":"Meyer","year":"1998","journal-title":"Pharm. Res."},{"issue":"11","key":"2026081909070729000_cit8","doi-asserted-by":"publisher","first-page":"4207","DOI":"10.1039\/C9NA00308H","article-title":"Hydrophobic\nIon Pairing: Encapsulating Small Molecules,\nPeptides, and Proteins into Nanocarriers","volume":"1","author":"Ristroph","year":"2019","journal-title":"Nanoscale\nAdv."},{"key":"2026081909070729000_cit9","doi-asserted-by":"publisher","first-page":"123507","DOI":"10.1016\/j.ijpharm.2023.123507","article-title":"Counterion Optimization\nfor Hydrophobic Ion Pairing (HIP): Unraveling the Key Factors","volume":"647","author":"Claus","year":"2023","journal-title":"Int. J. Pharm."},{"issue":"21","key":"2026081909070729000_cit10","doi-asserted-by":"publisher","first-page":"8515","DOI":"10.1039\/D4MA00720D","article-title":"Re-Engineering Lysozyme Solubility\nand Activity through Surfactant Complexation","volume":"5","author":"Mu","year":"2024","journal-title":"Mater. 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Protein Res."},{"issue":"1","key":"2026081909070729000_cit17","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1039\/B904761A","article-title":"Solubilisation of \u03b1-Chymotrypsin\nby Hydrophobic Ion Pairing in Fluorous Systems and Supercritical Carbon\nDioxide and Demonstration of Efficient Enzyme Recycling","volume":"12","author":"Benaissi","year":"2010","journal-title":"Green Chem."},{"issue":"1\u20132","key":"2026081909070729000_cit18","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/j.ijpharm.2010.03.036","article-title":"Biochemical and Biophysical Characterization\nof Lysozyme Modified by PEGylation","volume":"392","author":"da Sivla Freitas","year":"2010","journal-title":"Int. J.\nPharm."},{"issue":"6","key":"2026081909070729000_cit19","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1128\/jb.58.6.731-736.1949","article-title":"The Determination\nof Lysozyme","volume":"58","author":"Smolelis","year":"1949","journal-title":"J. Bacteriol."},{"key":"2026081909070729000_cit20","unstructured":"Gattefosse.\n          \n          Measuring saturation\nsolubility in lipid\nexcipients.; https:\/\/www.gattefosse.com\/characterization-methods\/measuring-saturation-solubility-lipid-excipients. (accessed 2025\u201305\u201328)."},{"issue":"1","key":"2026081909070729000_cit21","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1186\/s42649-020-00032-9","article-title":"Hot Stage Microscopy and Its Applications\nin Pharmaceutical Characterization","volume":"50","author":"Kumar","year":"2020","journal-title":"Appl. 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Rep."},{"issue":"12","key":"2026081909070729000_cit30","doi-asserted-by":"publisher","first-page":"3825","DOI":"10.3390\/ijms19123825","article-title":"The Role of Hydrogen\nBonding in the Folding\/Unfolding Process of Hydrated Lysozyme: A Review\nof Recent NMR and FTIR Results","volume":"19","author":"Mallamace","year":"2018","journal-title":"Int. J. Mol.\nSci."},{"issue":"1\u20132","key":"2026081909070729000_cit31","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1016\/j.ijpharm.2014.05.024","article-title":"Continuous Manufacturing\nof Solid Lipid Nanoparticles by Hot Melt Extrusion","volume":"471","author":"Patil","year":"2014","journal-title":"Int. J. 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Dairy Sci."},{"key":"2026081909070729000_cit35","doi-asserted-by":"publisher","first-page":"115318","DOI":"10.1016\/j.foodres.2024.115318","article-title":"Molten Globule-State\nProtein Structure: Perspectives from Food Processing Applications","volume":"198","author":"Lou","year":"2024","journal-title":"Food Res. Int."},{"key":"2026081909070729000_cit36","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1016\/j.ejpb.2016.10.021","article-title":"Effect of\nDrug Solubility and Lipid Carrier on Drug Release from Lipid Nanoparticles\nfor Dermal Delivery","volume":"110","author":"Zoubari","year":"2017","journal-title":"Eur. J. Pharm. Biopharm."},{"issue":"20","key":"2026081909070729000_cit37","doi-asserted-by":"publisher","first-page":"4610","DOI":"10.3390\/molecules25204610","article-title":"An EGF- and Curcumin-Co-Encapsulated\nNanostructured\nLipid Carrier Accelerates Chronic-Wound Healing in Diabetic Rats","volume":"25","author":"Lee","year":"2020","journal-title":"Molecules"},{"issue":"9","key":"2026081909070729000_cit38","doi-asserted-by":"publisher","first-page":"4664","DOI":"10.1021\/acs.molpharmaceut.3c00403","article-title":"Lysozyme\u2013Sucrose\nInteractions\nin the Solid State: Glass Transition, Denaturation, and the Effect\nof Residual Water","volume":"20","author":"Bogdanova","year":"2023","journal-title":"Mol. 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