Employing electrochemically derived pH gradients for Lab-on-PCB protein preconcentration implementations DOI Creative Commons
Grace Maxted, Pedro Estrela, Despina Moschou

et al.

Research Square (Research Square), Journal Year: 2023, Volume and Issue: unknown

Published: July 13, 2023

Abstract Protein preconcentration is an essential sample preparation step when analysing samples where the targeted proteins are in low concentrations, such as bodily fluids well water or wastewater. Nonetheless, very few practical implementations of miniaturized protein pre-concentration devices have been demonstrated practice and even fewer integration with other microanalytical steps. In this paper we propose for first time a isoelectric focusing-based protein-preconcentration device based on electrochemically derived pH gradients, rather than existing chemical reagent approaches. That way reducing need additional reagents to zero, whilst enabling incorporation seamlessly integrated full analysis microsystem via Lab-on-PCB technology. We apply our previously presented approach quantitatively control solution at vicinity planar electrodes using electrochemical generation acid through redox-active self-assembled monolayers. The was comprised printed circuit board array gold which functionalised 4-Aminothiophenol; formed monolayer electropolymerised improve its reversibility. performed two configurations, one open required use holder suspend fluid above electrodes, another used microfluidic channels enclose small volumes fluid. Reported here data both these forms quantitative concentration factor shown form qualitative proof microfluidic.

Language: Английский

Employing electrochemically derived pH gradients for Lab-on-PCB protein preconcentration devices DOI Creative Commons
Grace Maxted, Pedro Estrela, Despina Moschou

et al.

Microsystems & Nanoengineering, Journal Year: 2024, Volume and Issue: 10(1)

Published: Jan. 18, 2024

Protein preconcentration is an essential sample preparation step for analysis in which the targeted proteins exist low concentrations, such as bodily fluids, water, or wastewater. Nonetheless, very few practical implementations of miniaturized protein devices have been demonstrated practice, and even fewer integrated with other microanalytical steps. Existing approaches rely heavily on additional chemicals reagents introduce complexity to overall assay. In this paper, we propose a novel isoelectric focusing-based screening device based electrochemically derived pH gradients rather than existing chemical reagent approaches. way, reduce need zero while enabling incorporation seamlessly full microsystem via Lab-on-PCB technology. We apply our previously presented approach quantitatively control solution vicinity planar electrodes using electrochemical acid generation through redox-active self-assembled monolayers. The comprises printed circuit board array gold that were functionalized 4-aminothiophenol; formed monolayer was electropolymerized improve its reversibility. performed two configurations. first open needed use holder suspend well fluid above electrodes; second used microfluidic channels enclose small volumes fluid. Reported here are resulting data both these forms, quantitative concentration factor shown form qualitative proof microfluidic.

Language: Английский

Citations

1
Fluorescence Labeling of Peptides: Finding the Optimal Protocol for Coupling Various Dyes to ATCUN-like Structures DOI Creative Commons
Jordi C. J. Hintzen, Shitanshu Devrani, Andrew J. Carrod

et al.

ACS Organic & Inorganic Au, Journal Year: 2024, Volume and Issue: 4(5), P. 517 - 525

Published: June 4, 2024

Labeling of peptides and proteins with fluorescent dyes is a key step in functionalizing these structures for wide array biological assays. However, coupling strategies such have not been optimized the most common compounds, while this typically precious costly whole synthesis. We searched best conditions attachment widely used as 6-carboxyfluorescein, Rhodamine B, BODIPY-FL to peptides, where amino terminal Cu(II) Ni(II) binding site (ATCUN) were model system. Surprisingly, conventional methods dye proved be satisfactory yielded poor efficiency results. discovered that when labeling primary amines on uncommon synthesis activated pentafluorophenol (PFP) esters efficient strategy, expedited by microwave irradiation. Coupling secondary achieved efficiently through reagents HATU PyBOP. Furthermore, we employed our fluorescently labeled ATCUN studies sensing, showing changing fluorophore does significantly affect fluorescence quenching process discovering optimal linker length between core dye, expanding repertoire fluorophores can application.

Language: Английский

Citations

0
Employing electrochemically derived pH gradients for Lab-on-PCB protein preconcentration implementations DOI Creative Commons
Grace Maxted, Pedro Estrela, Despina Moschou

et al.

Research Square (Research Square), Journal Year: 2023, Volume and Issue: unknown

Published: July 13, 2023

Abstract Protein preconcentration is an essential sample preparation step when analysing samples where the targeted proteins are in low concentrations, such as bodily fluids well water or wastewater. Nonetheless, very few practical implementations of miniaturized protein pre-concentration devices have been demonstrated practice and even fewer integration with other microanalytical steps. In this paper we propose for first time a isoelectric focusing-based protein-preconcentration device based on electrochemically derived pH gradients, rather than existing chemical reagent approaches. That way reducing need additional reagents to zero, whilst enabling incorporation seamlessly integrated full analysis microsystem via Lab-on-PCB technology. We apply our previously presented approach quantitatively control solution at vicinity planar electrodes using electrochemical generation acid through redox-active self-assembled monolayers. The was comprised printed circuit board array gold which functionalised 4-Aminothiophenol; formed monolayer electropolymerised improve its reversibility. performed two configurations, one open required use holder suspend fluid above electrodes, another used microfluidic channels enclose small volumes fluid. Reported here data both these forms quantitative concentration factor shown form qualitative proof microfluidic.

Language: Английский

Citations

0