On-Chip Enrichment System for Digital Bioassay Based on Aqueous Two-Phase System.
ATPS
LLPS
digital bioassay
enrichment
microreactors
Journal
ACS nano
ISSN: 1936-086X
Titre abrégé: ACS Nano
Pays: United States
ID NLM: 101313589
Informations de publication
Date de publication:
10 01 2023
10 01 2023
Historique:
pubmed:
30
12
2022
medline:
12
1
2023
entrez:
29
12
2022
Statut:
ppublish
Résumé
We developed an on-chip enrichment method based on an aqueous two-phase system of dextran/polyethylene glycol mix, DEX/PEG ATPS, for digital bioassay. Accordingly, we prepared an array device of femtoliter reactors that displays millions of uniformly shaped DEX-rich droplets under a PEG-rich medium. The DEX-rich droplets effectively enriched DNA molecules from the PEG-rich medium. To quantify the enrichment power of the system, we performed a digital bioassay of alkaline phosphatase (ALP). Upon genetically tagging ALP molecules with the DEX-binding domain (DBD) derived from dextransucrase, the ALP molecules were enriched 59-fold in the DEX droplets in comparison to that in a conventional digital bioassay. Subsequently, we performed a Cas13-based digital SARS-CoV-2 RNA detection assay to evaluate the performance of this system for a more practical assay. In this assay, the target RNA molecules bound to the DBD-tagged Cas13 molecules were effectively enriched in the DEX droplets. Consequently, an enrichment factor of 31 was achieved. Enrichment experiments for nonlabeled proteins were also performed to test the expandability of this technique. The model protein, nontagged β-galactosidase, was enriched in DEX droplets containing DBD-tagged antibody, with an enrichment factor of over 100. Thus, this system enabled effective on-chip enrichment of target molecules to enhance the detection sensitivity of digital bioassays without using external instruments or an external power source, which would be applicable for on-site bioassays or portable diagnostic tests.
Identifiants
pubmed: 36579744
doi: 10.1021/acsnano.2c06007
pmc: PMC9835982
doi:
Substances chimiques
Polyethylene Glycols
3WJQ0SDW1A
RNA, Viral
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
212-220Références
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