Integrated Extraction-Preservation Strategies for RNA Using Biobased Ionic Liquids
authors Quental, MV; Pedro, AQ; Pereira, P; Sharma, M; Queiroz, JA; Coutinho, JAP; Sousa, F; Freire, MG
nationality International
journal ACS SUSTAINABLE CHEMISTRY & ENGINEERING
author keywords Ribonucleic acid; Amino-acid-based ionic liquids; Aqueous biphasic systems; Integrated extraction preservation process
keywords AQUEOUS BIPHASIC SYSTEMS; SINGLE-STEP METHOD; SECONDARY STRUCTURES; PROTEIN SEPARATION; 2-PHASE SYSTEMS; GOODS BUFFERS; ACID; PURIFICATION; STABILITY; CHROMATOGRAPHY
abstract The ubiquitous instability of RNA along with issues associated with its purity degree have been preventing its widespread use as low-cost biotherapeutics. On the basis of the well-known capacity of amino acids to specifically interact with RNA when used as chromatographic ligands, a set of amino-acid-based ionic liquids (AA-ILs) was herein investigated, both to act as preservation media and as phase forming agents of aqueous biphasic systems (ABS). This set of strategies was combined with the goal of developing integrated extraction-preservation platforms. AA-ILs comprising the cholinium cation and anions derived from L-lysine ([Ch][Lys]), L-arginine ([Ch][Arg]), L-glutamic acid ([Ch][Glu]), and DL-aspartic acid ([Ch] [Asp]) were studied. It is shown that the stability of RNA is preserved in aqueous solutions of the studied AA-ILs, even in the presence of ribonucleases (RNases). Furthermore, almost all the investigated AA-ILs display no cytotoxicity onto two distinct human cell lines. After identifying the most promising ILs, ABS formed by AA-ILs and polypropylene glycol with a molecular weight of 400 g morl (PPG 400) were investigated as extraction and purification platforms for RNA. Both with pure RNA and bacterial lysate samples, RNA is successfully extracted to the IL-rich phase without compromising its integrity and stability. On the basis of these results, the integrated extraction-preservation process for RNA is finally demonstrated. RNA is initially extracted from the bacterial lysate sample using ABS, after which the IL-rich phase can be used as the preservation medium of RNA up to its use. RNA can be then recovered from the IL-rich phase by ethanol precipitation, and the ABS phase-forming components recovered and reused. Although improvements in the purity level of RNA are still required, the approach here reported represents a step forward in the development of sustainable processes to overcome the critical demand of high-quality/high-purity RNA to be used as biotherapeutics.
publisher AMER CHEMICAL SOC
issn 2168-0485
year published 2019
volume 7
issue 10
beginning page 9439
ending page 9448
digital object identifier (doi) 10.1021/acssuschemeng.9b00688
web of science category Chemistry, Multidisciplinary; Green & Sustainable Science & Technology; Engineering, Chemical
subject category Chemistry; Science & Technology - Other Topics; Engineering
unique article identifier WOS:000469304900041

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