{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:49:59Z","timestamp":1760143799154,"version":"build-2065373602"},"reference-count":110,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,2,27]],"date-time":"2024-02-27T00:00:00Z","timestamp":1708992000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"FCT\/MCTES (Portugal)","doi-asserted-by":"publisher","award":["PTDC\/EQU-EQU\/2223\/2021"],"award-info":[{"award-number":["PTDC\/EQU-EQU\/2223\/2021"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Interferon alpha-2b (IFN-\u03b12b) is an essential cytokine widely used in the treatment of chronic hepatitis C and hairy cell leukemia, and serum albumin is the most abundant plasma protein with numerous physiological functions. Effective single-step aqueous biphasic system (ABS) extraction for the simultaneous purification of IFN-\u03b12b and BSA (serum albumin protein) was developed in this work. Effects of the ionic liquid (IL)-based ABS functionalization, fluorinated ILs (FILs; [C\u200b2C\u200b1Im][C\u200b4F\u200b9SO\u200b3] and [N\u200b1112(OH)][C\u200b4F\u200b9SO\u200b3]) vs. mere fluoro-containing IL ([C\u200b4C\u200b1Im][CF\u200b3SO\u200b3]), in combination with sucrose or [N\u200b1112(OH)][H\u200b2PO\u200b4] (well-known globular protein stabilizers), or high-charge-density salt K\u200b3PO\u200b4 were investigated. The effects of phase pH, phase water content (%wt), phase composition (%wt), and phase volume ratio were investigated. The phase pH was found to have a significant effect on IFN-\u03b12b and BSA partition. Experimental results show that simultaneous single-step purification was achieved with a high yield (extraction efficiency up to 100%) for both proteins and a purification factor of IFN-\u03b12b high in the enriched IFN-\u03b12b phase (up to 23.22) and low in the BSA-enriched phase (down to 0.00). SDS-PAGE analysis confirmed the purity of both recovered proteins. The stability and structure of IFN-\u03b12b and BSA were preserved or even improved (FIL-rich phase) during the purification step, as evaluated by CD spectroscopy and DSC. Binding studies of IFN-\u03b12b and BSA with the ABS phase-forming components were assessed by MST, showing the strong interaction between FILs aggregates and both proteins. In view of their biocompatibility, customizable properties, and selectivity, FIL-based ABSs are suggested as an improved purification step that could facilitate the development of biologics.<\/jats:p>","DOI":"10.3390\/ijms25052751","type":"journal-article","created":{"date-parts":[[2024,2,27]],"date-time":"2024-02-27T11:52:55Z","timestamp":1709034775000},"page":"2751","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Simultaneous Purification of Human Interferon Alpha-2b and Serum Albumin Using Bioprivileged Fluorinated Ionic Liquid-Based Aqueous Biphasic Systems"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5751-9933","authenticated-orcid":false,"given":"Sara F.","family":"Carvalho","sequence":"first","affiliation":[{"name":"LAQV, REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, NOVA University Lisbon, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7166-6764","authenticated-orcid":false,"given":"Ana B.","family":"Pereiro","sequence":"additional","affiliation":[{"name":"LAQV, REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, NOVA University Lisbon, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8648-7539","authenticated-orcid":false,"given":"Jo\u00e3o M. M.","family":"Ara\u00fajo","sequence":"additional","affiliation":[{"name":"LAQV, REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, NOVA University Lisbon, 2829-516 Caparica, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"873","DOI":"10.2165\/00003495-199550050-00007","article-title":"Interferon-\u03b1-2a.A Review of its Pharmacological Properties and Therapeutic Use in the Management of Viral Hepatitis","volume":"50","author":"Haria","year":"1995","journal-title":"Drugs"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1016\/S1569-2582(97)80010-4","article-title":"Chapter 26 the interferons","volume":"9","author":"Baron","year":"1998","journal-title":"Princ. Med Biol."},{"key":"ref_3","first-page":"258","article-title":"Virus interference. I. The interferon","volume":"147","author":"Isaacs","year":"1957","journal-title":"Proc. R. Soc. 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