Multimodal ionic liquid-based chromatographic supports for an effective RNA purification

resumo

Nucleic acids have been considered interesting molecules to be used as biopharmaceuticals for the treatment of various diseases, in gene therapy strategies. In particular, RNA arises as the most promising approach because it does not require access to the nucleus of cells to exert its function; however, it is quite challenging due to its labile nature. To increase the possibility of translating RNA-based technology to clinical protocols, the biomanufacturing of RNAs has been intensively exploited in the last few years. However, the standard RNA purification processes remain time-consuming and present limitations regarding recovery yield and purity. This work describes the functionalization of chromatographic silica-based supports with four ionic liquids (ILs) composed of functional moieties that can promote distinct interactions with nucleic acids. After an initial screening to evaluate the binding and elution behavior of nucleic acids in the IL-based supports, SSi[C(3)C(3NH2)Im]Cl has shown to be the most promising for further purification assays. This support was studied for the RNA purification from different samples (clarified or more complex) and has shown to be highly effective, for all the conditions studied. Generally, it is here presented a new method for RNA isolation in a single step, using an IL-based chromatographic support, able to eliminate the usage of hazardous compounds often included in standard RNA extraction protocols.

palavras-chave

STATIONARY-PHASE; SEPARATION; EXTRACTION; DNA

categoria

Engineering

autores

Carapito, R; Bernardo, SC; Pereira, MM; Neves, MC; Freire, MG; Sousa, F

Grupos

G5 - Materiais Biomiméticos, Biológicos e Vivos
G6 - Materiais Virtuais e Inteligência Artificial

Projectos

CICECO - Aveiro Institute of Materials (UIDB/50011/2020)

CICECO - Aveiro Institute of Materials (UIDP/50011/2020)

Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)

Yeast cell factory for mRNA bioproduction (Yscript)

Rede Nacional de Ressonância Magnética Nuclear (PTNMR)

agradecimentos

The authors acknowledge the funding from Portuguese Foundation for Science and Technology (FCT) , through the projects UIDB/00709/2020, UIDP/00709/2020 and the project PTDC/BII-BBF/29496/2017 funded by FEDER, through COMPETE2020-POCI, and by national funds, through FCT/MCTES. This work was also developed within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020, UIDP/50011/2020 & LA/P/0006/2020, financed by national funds through the FCT/MCTES (PIDDAC). CIEPQPF is supported by the FCT through the projects UIDB/EQU/00102/2020 and UIDP/EQU/00102/2020. The work was developed within the scope of the EIC-Pathfinder YSCRIPT project with reference 101047. The NMR spectrometers are part of the National NMR Network (PTNMR) and are partially supported by Infrastructure Project No 022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC) . Rita Carapito and Marcia C. Neves acknowledge FCT for the PhD fellowship (2020.07980.BD) and the research contract CEECIND/00383/2017, respectively.