Cholinium-Based Good's Buffers Ionic Liquids as Remarkable Stabilizers and Recyclable Preservation Media for Recombinant Small RNAs
authors Pedro, AQ; Pereira, P; Quental, MJ; Carvalho, AP; Santos, SM; Queiroz, JA; Sousa, F; Freire, MG
nationality International
author keywords Nucleic acids; Recombinant RNA; Ionic liquids; Good's buffers; Stability; Recyclable media
abstract RNA is a biopolymer of high relevance in the biopharmaceuticals field and in fundamental and applied research; however, preservation of RNA stability is still a remarkable challenge. Herein, we demonstrate the enhanced potential of aqueous solutions of self-buffering cholinium-based Good's buffers ionic liquids (GB-ILs), at 20 and 50% (w/w), as alternative preservation media of recombinant small RNAs. The thermal stability of RNA is highly enhanced by GB-ILs, with an increase of 14 degrees C in the biopolymer melting temperature-the highest increase observed to date with ILs. Most GB-ILs investigated improve the stability of RNA at least up to 30 days, both at 25 degrees C and at 4 degrees C, without requiring the typical samples freezing. Molecular dynamics simulations were applied to better understand the molecular-level mechanisms responsible for the observed RNA improved stability. The number of IL cations surrounding the RNA chain is similar, yet with differences found for the IL anions, which are responsible for the overall charge of the biopolymer first solvation sphere. No cytotoxicity of the studied solutions containing RNA and ILs at 20% (w/w) was observed onto two distinct human cell lines, reinforcing their potential to act as preservation media when foreseeing biopharmaceutical applications. Finally, RNA was successfully recovered from the ILs aqueous solutions, without changes in its structural integrity, and the ILs were successfully recycled and reused.
issn 2168-0485
year published 2018
volume 6
issue 12
beginning page 16645
ending page 16656
digital object identifier (doi) 10.1021/acssuschemeng.8b03900
web of science category Chemistry, Multidisciplinary; Green & Sustainable Science & Technology; Engineering, Chemical
subject category Chemistry; Science & Technology - Other Topics; Engineering
unique article identifier WOS:000452344900077
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