Simultaneous Purification of Human Interferon Alpha-2b and Serum Albumin Using Bioprivileged Fluorinated Ionic Liquid-Based Aqueous Biphasic Systems.
Int J Mol Sci
; 25(5)2024 Feb 27.
Article
em En
| MEDLINE
| ID: mdl-38473998
ABSTRACT
Interferon alpha-2b (IFN-α2b) 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-α2b and BSA (serum albumin protein) was developed in this work. Effects of the ionic liquid (IL)-based ABS functionalization, fluorinated ILs (FILs; [Câ2Câ1Im][Câ4Fâ9SOâ3] and [Nâ1112(OH)][Câ4Fâ9SOâ3]) vs. mere fluoro-containing IL ([Câ4Câ1Im][CFâ3SOâ3]), in combination with sucrose or [Nâ1112(OH)][Hâ2POâ4] (well-known globular protein stabilizers), or high-charge-density salt Kâ3POâ4 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-α2b 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-α2b high in the enriched IFN-α2b 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-α2b 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-α2b 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.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Albumina Sérica
/
Líquidos Iônicos
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
Portugal