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An ATPS‐based droplet microfluidic device for protein partitioning.

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Title: An ATPS‐based droplet microfluidic device for protein partitioning.
Authors: Moreno‐Sibaja, Federico1 (AUTHOR), Hernández‐Cid, David2 (AUTHOR), Ceballos‐Medina, Javier3,4 (AUTHOR), Cerón‐Torres, Daniel Abirán5 (AUTHOR), Reyes‐Avendaño, Jorge Antonio5 (AUTHOR), Gallo‐Villanueva, Roberto Carlos5 (AUTHOR), Pérez‐Gonzalez, Victor Hugo5 (AUTHOR), Mata‐Gómez, Marco Arnulfo5 (AUTHOR) mmatag@tec.mx
Source: Journal of Chemical Technology & Biotechnology. Jul2026, Vol. 101 Issue 7, p1324-1331. 8p.
Subjects: Microfluidic devices, Protein fractionation, Microencapsulation, Fluorescent proteins, Polyethylene glycol, Liquid-liquid extraction, Microfluidics
Abstract: Background: Droplet microfluidics uses immiscible phases to manipulate discrete volumes inside microchannels, and it is applied in synthesis of biomolecules, encapsulation, drug delivery and cell analysis. The use of aqueous two‐phase systems (ATPS) is a strategy for the fractionation of biomolecules. Here, we combined both techniques to fabricate a microfluidic device based on a polymer–salt ATPS, composed of polyethylene glycol (PEG) 8000 and phosphate salt buffer (PO4), capable of generating droplets and partitioning red fluorescent protein (RFP), used as a model of study. Partition behavior, droplet size and frequency were characterized, employing a polydimethylsiloxane microdevice for this purpose. Results: The microdevice exhibited either a two‐phase or droplet behavior in the absence and presence of RFP. In both scenarios, the model protein was partitioned towards the salt‐rich phase (PO4), as in traditional ATPS. In addition, by controlling fluid velocity, different regimes of droplet formation were observed, i.e. dripping and jetting. RFP was partitioned and encapsulated in both regimes. The microdevices exhibited a partition coefficient, Kp, of 0.34 ± 0.016, which is comparable with that obtained in traditional systems (Kp = 0.39 ± 0.012). Droplet size diminished as Ca, μU/σ, increased. Droplet breakup time has the same effect when increasing velocity ratio, while frequency of droplet formation increased. COMSOL computational simulations were performed, exhibiting good agreement with experimental results. Conclusion: The microfluidic device proved to be a useful platform to generate droplets by using polymer PEG–salt ATPS under controlled conditions. Besides, it allowed partitioning and encapsulation of the model protein. This microfluidic platform presents a viable alternative for protein separation and encapsulation, which can further support high‐value‐biomolecule separation. © 2025 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]
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Database: Engineering Source
Description
Abstract:Background: Droplet microfluidics uses immiscible phases to manipulate discrete volumes inside microchannels, and it is applied in synthesis of biomolecules, encapsulation, drug delivery and cell analysis. The use of aqueous two‐phase systems (ATPS) is a strategy for the fractionation of biomolecules. Here, we combined both techniques to fabricate a microfluidic device based on a polymer–salt ATPS, composed of polyethylene glycol (PEG) 8000 and phosphate salt buffer (PO4), capable of generating droplets and partitioning red fluorescent protein (RFP), used as a model of study. Partition behavior, droplet size and frequency were characterized, employing a polydimethylsiloxane microdevice for this purpose. Results: The microdevice exhibited either a two‐phase or droplet behavior in the absence and presence of RFP. In both scenarios, the model protein was partitioned towards the salt‐rich phase (PO4), as in traditional ATPS. In addition, by controlling fluid velocity, different regimes of droplet formation were observed, i.e. dripping and jetting. RFP was partitioned and encapsulated in both regimes. The microdevices exhibited a partition coefficient, Kp, of 0.34 ± 0.016, which is comparable with that obtained in traditional systems (Kp = 0.39 ± 0.012). Droplet size diminished as Ca, μU/σ, increased. Droplet breakup time has the same effect when increasing velocity ratio, while frequency of droplet formation increased. COMSOL computational simulations were performed, exhibiting good agreement with experimental results. Conclusion: The microfluidic device proved to be a useful platform to generate droplets by using polymer PEG–salt ATPS under controlled conditions. Besides, it allowed partitioning and encapsulation of the model protein. This microfluidic platform presents a viable alternative for protein separation and encapsulation, which can further support high‐value‐biomolecule separation. © 2025 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]
ISSN:02682575
DOI:10.1002/jctb.7816