resumo
Nucleic acid-based treatments have become increasingly important in addressing diseases linked to genetic defects. In Alzheimer's disease, decreased levels of microRNA-29b lead to the overproduction of the human betasecretase 1 (hBACE1) enzyme, which is associated with the progression of the condition. Consequently, pre-miR29b represents a promising therapeutic agent, as restoring its levels could diminish hBACE1 expression and reduce amyloid-beta accumulation in neuronal cells. However, the successful deployment of pre-miRNA-29b as a treatment relies heavily on ensuring its purity, stability, and biological potency. This work evaluates the selective purification of pre-miRNA-29b, recombinantly produced from Rhodovulum sulfidophilum, using Capto Q ImpRes, a multi-modal strong anion exchange resin. The research optimizes process parameters, such as NaCl concentration and pH, to maximize the recovery and purity of pre-miRNA-29b through a Central Composite Design (CCD). The results reveal a relationship between recovery and purity, with CCD predicting optimal conditions yielding up to 48.21% recovery and 51.15% purity of pre-miRNA-29b. Overall, this work highlights the potential of using multimodal resins to selectively capture pre-miRNA-29b from complex recombinant samples. This selectivity demonstrates the promise of this technique for the isolation of specific RNA molecules, a crucial step in the development of effective nucleic acid-based therapies. These findings contribute to the ongoing efforts to establish highly effective purification strategies that ensure the quality of therapeutic RNAs.
palavras-chave
RESINS
categoria
Engineering
autores
Carapito, R; Bento, J; Sponchioni, M; Freire, MG; Sousa, F
nossos autores
Mara Guadalupe Freire Martins
Professor CatedráticoProjectos
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)
agradecimentos
This work was developed within the scope of the projects of RISE-Health, UBI: UIDB/00709/2020 (DOI 10.54499/UIDB/00709/2020) and UIDP/00709/2020 (DOI 10.54499/UIDP/00709/2020), of CICECO-Aveiro Institute of Materials: UIDB/50011/2020 (DOI 10.54499/UIDB/50011/2020), UIDP/50011/2020 (DOI 10.54499/UIDP/50011/2020) & LA/P/0006/2020 (DOI 10.54499/LA/P/0006/2020), financed by national funds through the FCT/MCTES (PIDDAC). Thanks are also due to the project PureDNA-Ref. 2022.03394.PTDC (http://doi.org/10.54499/2022.03394.PTDC), financially supported by national funds (OE), through FCT/MCTES. Rita Carapito also acknowledges FCT for the Ph.D. fellowship (DOI 10.54499/2020.07980.BD).