Integrated platform resorting to ionic liquids comprising the extraction, purification and preservation of DNA

[Display omitted] •Three phase partitioning resorting to aqueous biphasic systems (ABS/TPP) were investigated.•A series of cholinium-based ionic liquids and different polymers were investigated to create ABS/TPP.•Designed ABS/TPP allow the simultaneous separation of dsDNA from DNase I.•The systems comprising cholinium glycolate is the best to simultaneously separate dsDNA from DNase I.•dsDNA enriches in the ionic-liquid-rich phase whereas DNase I precipitates at the interphase. The large-scale production of therapeutically targeted-deoxyribonucleic acid (DNA) has passed through several challenges, postponing the tangible implementation of an effective, economic and sustainable manufacturing system. Such challenges comprise the need to develop an integrative downstream process able to extract, purify and long-term preserve DNA, whilst reducing the risk of degradation by endonucleases that would compromise their effectiveness as therapeutic products. In this work, three-phase partitioning (TPP) systems formed by the application of aqueous biphasic systems (ABS) composed of several biocompatible cholinium-based ionic liquids (ILs), are proposed for the separation of double stranded DNA (dsDNA) from the endonuclease deoxyribonuclease I (DNase I). By taking advantage of the tailor-made properties of ILs, dsDNA can be completely extracted to the IL-rich phase, whereas DNase I is precipitated at the ABS interphase. The ABS/TPP formed by IL cholinium glycolate ([N111(2OH)][Gly]) fulfills the aim of this work, i.e. at ensuring the technical viability of IL-based ABS/TPP for the “one-pot” extraction, purification and long-term preservation of dsDNA. The results reveal the potential of this system to be applied in the bioprocessing of DNA, particularly relevant when envisioning DNA-based therapeutic products.