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Main Authors: Sakuta, Hiroki, Akamine, Yuki, Kamo, Akari, Gong, Hao, Ichihashi, Norikazu, Nikoubashman, Arash, Yanagisawa, Miho
Format: Preprint
Published: 2025
Subjects:
Online Access:https://arxiv.org/abs/2509.24641
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author Sakuta, Hiroki
Akamine, Yuki
Kamo, Akari
Gong, Hao
Ichihashi, Norikazu
Nikoubashman, Arash
Yanagisawa, Miho
author_facet Sakuta, Hiroki
Akamine, Yuki
Kamo, Akari
Gong, Hao
Ichihashi, Norikazu
Nikoubashman, Arash
Yanagisawa, Miho
contents Mixtures of polyethylene glycol (PEG) and dextran (Dex) represent a widely used class of aqueous two-phase systems (ATPS), with applications ranging from the purification of various biomolecules such as nucleic acids to the synthesis of protocells. A key feature underlying these applications is the selective accumulation of biomolecules within Dex-rich droplets in an aqueous PEG phase, but the physical origin of this partitioning remains unclear. Entropic interactions were long assumed to be the primary driving force; however, our systematic experiments using DNA of different lengths indicate that entropy alone cannot fully explain the observed behavior. We identify an additional and previously underappreciated contribution from electrostatic interactions: Dex carries a slightly more negative charge than PEG, which drives preferential cation accumulation in the Dex-rich phase. These counterions facilitate the selective partitioning of DNA inside the Dex-rich droplets. This mechanism explains the dependency of DNA uptake in Dex-rich droplets on polymer length and salt concentration. Our findings establish that Donnan-type ion partitioning plays a crucial role in the localization of long nucleic acids in Dex-rich droplets, offering a unified explanation for this long-standing phenomenon. They lay the foundation for designing ATPS-based systems and help elucidate the physicochemical principles of biomolecular partition upon phase separation in cells.
format Preprint
id arxiv_https___arxiv_org_abs_2509_24641
institution arXiv
publishDate 2025
record_format arxiv
spellingShingle Cation accumulation drives the preferential partitioning of DNA in an aqueous two-phase system
Sakuta, Hiroki
Akamine, Yuki
Kamo, Akari
Gong, Hao
Ichihashi, Norikazu
Nikoubashman, Arash
Yanagisawa, Miho
Soft Condensed Matter
Mixtures of polyethylene glycol (PEG) and dextran (Dex) represent a widely used class of aqueous two-phase systems (ATPS), with applications ranging from the purification of various biomolecules such as nucleic acids to the synthesis of protocells. A key feature underlying these applications is the selective accumulation of biomolecules within Dex-rich droplets in an aqueous PEG phase, but the physical origin of this partitioning remains unclear. Entropic interactions were long assumed to be the primary driving force; however, our systematic experiments using DNA of different lengths indicate that entropy alone cannot fully explain the observed behavior. We identify an additional and previously underappreciated contribution from electrostatic interactions: Dex carries a slightly more negative charge than PEG, which drives preferential cation accumulation in the Dex-rich phase. These counterions facilitate the selective partitioning of DNA inside the Dex-rich droplets. This mechanism explains the dependency of DNA uptake in Dex-rich droplets on polymer length and salt concentration. Our findings establish that Donnan-type ion partitioning plays a crucial role in the localization of long nucleic acids in Dex-rich droplets, offering a unified explanation for this long-standing phenomenon. They lay the foundation for designing ATPS-based systems and help elucidate the physicochemical principles of biomolecular partition upon phase separation in cells.
title Cation accumulation drives the preferential partitioning of DNA in an aqueous two-phase system
topic Soft Condensed Matter
url https://arxiv.org/abs/2509.24641