File Download
There are no files associated with this item.
Links for fulltext
(May Require Subscription)
- Publisher Website: 10.1021/acs.analchem.2c04861
- Scopus: eid_2-s2.0-85149122725
- Find via
Supplementary
-
Citations:
- Scopus: 0
- Appears in Collections:
Article: Partitioning-Induced Isolation of Analyte and Analysis via Multiscaled Aqueous Two-Phase System
Title | Partitioning-Induced Isolation of Analyte and Analysis via Multiscaled Aqueous Two-Phase System |
---|---|
Authors | |
Issue Date | 14-Mar-2023 |
Publisher | American Chemical Society |
Citation | Analytical Chemistry, 2023, v. 95, n. 10, p. 4644-4652 How to Cite? |
Abstract | Most fluorescence-based bioanalytical applications need labeling of analytes. Conventional labeling requires washing to remove the excess fluorescent labels and reduce the noise signals. These pretreatments are labor intensive and need specialized equipment, hindering portable applications in resource-limited areas. Herein, we use the aqueous two-phase system (ATPS) to realize the partitioning-induced isolation of labeled analytes from background signals without extra processing steps. ATPS is formed by mixing two polymers at sufficiently high concentrations. ATPS-based isolation is driven by intrinsic affinity differences between analytes and excess labels. To demonstrate the partitioning-induced isolation and analysis, fluorescein isothiocyanate (FITC) is selected as the interfering fluorophore, and a monoclonal antibody (IgG) is used as the analyte. To optimize ATPS compositions, different molecular weights and mass fractions of polyethylene glycol (PEG) and dextran and different phosphate-buffered saline (PBS) concentrations are investigated. Various operational scales of our approach are demonstrated, suggesting its compatibility with various bioanalytical applications. In centimeter-scale ATPS, the optimized distribution ratios of IgG and FITC are 91.682 and 0.998 using PEG 6000 Da and dextran 10,000 Da in 10 mM PBS. In millimeter-scale ATPS, the analyte is enriched to 6.067 fold using 15 wt % PEG 35,000 Da and 5 wt % dextran 500,000 Da in 10 mM PBS. In microscale ATPS, analyte dilutions are isolated into picoliter droplets, and the measured fluorescence intensities linearly correlated with the analyte concentrations (R2 = 0.982). |
Persistent Identifier | http://hdl.handle.net/10722/338543 |
ISSN | 2021 Impact Factor: 8.008 2020 SCImago Journal Rankings: 2.117 |
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Cao, Y | - |
dc.contributor.author | Tian, J | - |
dc.contributor.author | Lin, H | - |
dc.contributor.author | Li, Q | - |
dc.contributor.author | Xiao, Y | - |
dc.contributor.author | Cui, H | - |
dc.contributor.author | Shum, HC | - |
dc.date.accessioned | 2024-03-11T10:29:41Z | - |
dc.date.available | 2024-03-11T10:29:41Z | - |
dc.date.issued | 2023-03-14 | - |
dc.identifier.citation | Analytical Chemistry, 2023, v. 95, n. 10, p. 4644-4652 | - |
dc.identifier.issn | 0003-2700 | - |
dc.identifier.uri | http://hdl.handle.net/10722/338543 | - |
dc.description.abstract | <p>Most fluorescence-based bioanalytical applications need labeling of analytes. Conventional labeling requires washing to remove the excess fluorescent labels and reduce the noise signals. These pretreatments are labor intensive and need specialized equipment, hindering portable applications in resource-limited areas. Herein, we use the aqueous two-phase system (ATPS) to realize the partitioning-induced isolation of labeled analytes from background signals without extra processing steps. ATPS is formed by mixing two polymers at sufficiently high concentrations. ATPS-based isolation is driven by intrinsic affinity differences between analytes and excess labels. To demonstrate the partitioning-induced isolation and analysis, fluorescein isothiocyanate (FITC) is selected as the interfering fluorophore, and a monoclonal antibody (IgG) is used as the analyte. To optimize ATPS compositions, different molecular weights and mass fractions of polyethylene glycol (PEG) and dextran and different phosphate-buffered saline (PBS) concentrations are investigated. Various operational scales of our approach are demonstrated, suggesting its compatibility with various bioanalytical applications. In centimeter-scale ATPS, the optimized distribution ratios of IgG and FITC are 91.682 and 0.998 using PEG 6000 Da and dextran 10,000 Da in 10 mM PBS. In millimeter-scale ATPS, the analyte is enriched to 6.067 fold using 15 wt % PEG 35,000 Da and 5 wt % dextran 500,000 Da in 10 mM PBS. In microscale ATPS, analyte dilutions are isolated into picoliter droplets, and the measured fluorescence intensities linearly correlated with the analyte concentrations (R2 = 0.982).</p> | - |
dc.language | eng | - |
dc.publisher | American Chemical Society | - |
dc.relation.ispartof | Analytical Chemistry | - |
dc.rights | This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. | - |
dc.title | Partitioning-Induced Isolation of Analyte and Analysis via Multiscaled Aqueous Two-Phase System | - |
dc.type | Article | - |
dc.identifier.doi | 10.1021/acs.analchem.2c04861 | - |
dc.identifier.scopus | eid_2-s2.0-85149122725 | - |
dc.identifier.volume | 95 | - |
dc.identifier.issue | 10 | - |
dc.identifier.spage | 4644 | - |
dc.identifier.epage | 4652 | - |
dc.identifier.eissn | 1520-6882 | - |
dc.identifier.issnl | 0003-2700 | - |