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Protocell Arrays for Simultaneous Detection of Diverse Analytes

View ORCID ProfileYan Zhang, View ORCID ProfileTaisuke Kojima, Ge-Ah Kim, View ORCID ProfileMonica P. McNerney, View ORCID ProfileShuichi Takayama, View ORCID ProfileMark P. Styczynski
doi: https://doi.org/10.1101/2021.02.13.431022
Yan Zhang
1School of Chemical & Biomolecular Engineering, Georgia Institute of Technology
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Taisuke Kojima
2Department of Biomedical Engineering, Georgia Institute of Technology
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Ge-Ah Kim
3School of Materials Science and Engineering, Georgia Institute of Technology
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Monica P. McNerney
1School of Chemical & Biomolecular Engineering, Georgia Institute of Technology
4Current address: Department of Systems Biology, Harvard Medical School
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Shuichi Takayama
2Department of Biomedical Engineering, Georgia Institute of Technology
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  • For correspondence: mark.styczynski@chbe.gatech.edu
Mark P. Styczynski
1School of Chemical & Biomolecular Engineering, Georgia Institute of Technology
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  • For correspondence: mark.styczynski@chbe.gatech.edu
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Abstract

Simultaneous detection of multiple analytes from a single sample (multiplexing), particularly when at the point of need, can guide complex decision-making without increasing the required sample volume or cost per test. Despite recent advances, multiplexing still typically faces the critical limitation of measuring only one type of molecule per assay platform – for example, only small molecules or only nucleic acids. In this work, we address this bottleneck with a customizable platform that integrates cell-free expression (CFE) with a polymer-based aqueous two-phase system (ATPS) to produce membrane-less “protocells” containing transcription and translation machinery used for analyte detection. Multiple protocells are arrayed in microwells where each protocell droplet performs distinct reactions to detect chemically diverse targets including small molecules, minerals, and nucleic acid sequences, all from the same sample. We demonstrate that these protocell arrays can measure analytes in a human biofluid matrix, maintain function after lyophilization and rehydration, and produce visually interpretable readouts, illustrating its potential for application as a minimal-equipment, field-deployable, multi-analyte detection tool.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted February 13, 2021.
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Protocell Arrays for Simultaneous Detection of Diverse Analytes
Yan Zhang, Taisuke Kojima, Ge-Ah Kim, Monica P. McNerney, Shuichi Takayama, Mark P. Styczynski
bioRxiv 2021.02.13.431022; doi: https://doi.org/10.1101/2021.02.13.431022
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Protocell Arrays for Simultaneous Detection of Diverse Analytes
Yan Zhang, Taisuke Kojima, Ge-Ah Kim, Monica P. McNerney, Shuichi Takayama, Mark P. Styczynski
bioRxiv 2021.02.13.431022; doi: https://doi.org/10.1101/2021.02.13.431022

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