An on-chip fluorogenic enzyme assay using a multilayer microchip interconnected with a nanocapillary array membrane

Maojun Gong; Bo Young Kim; Bruce R. Flachsbart; Mark A. Shannon; Paul W. Bohn; Jonathan V. Sweedler

doi:10.1109/JSEN.2008.918912

An on-chip fluorogenic enzyme assay using a multilayer microchip interconnected with a nanocapillary array membrane

Maojun Gong, Bo Young Kim, Bruce R. Flachsbart, Mark A. Shannon, Paul W. Bohn, Jonathan V. Sweedler

Research output: Contribution to journal › Article › peer-review

Abstract

Microfluidic devices allow manipulation of reagents and fluids in a semi-automated fashion, ideal for performing multiple measurements or conditioning various reagents. Here, an enzyme assay has been performed in a multilayer poly(methyl methacrylate)-based microfluidic device, where the layers are fluidically connected via a nanocapillary array membrane serving as an effective injector and valve. As a model system, β-glucuronidase from Escherichia coli and fluorescein di(β-D-glucuronide) are used for the assay; offline mixing and online incubation of substrate and enzyme allow determination of the initial hydrolysis rates of the substrate under catalysis by β-glucuronidase. The Michaelis constant K_m was determined to be ∼ 4.0 μM for the enzyme of 83 units/mL at ambient temperature. The 50% inhibitory concentration IC₅₀ of D-saccharic acid-1,4-Iactone to 167 units/mL was estimated to be 3.0 μtM. These results demonstrate added functionality for a poly(methyl methacrylate)-based nanocapillary array membrane-containing microfluidic device for following enzyme reaction kinetics.

Original language	English (US)
Article number	4529140
Pages (from-to)	601-607
Number of pages	7
Journal	IEEE Sensors Journal
Volume	8
Issue number	5
DOIs	https://doi.org/10.1109/JSEN.2008.918912
State	Published - May 2008

Keywords

Enzyme kinetics
Microcontact printing
Microfluidics
Nanocapillary array membranes
Nanofluidics
Poly(methyl methacrylate)

ASJC Scopus subject areas

Engineering(all)
Electrical and Electronic Engineering

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title = "An on-chip fluorogenic enzyme assay using a multilayer microchip interconnected with a nanocapillary array membrane",

abstract = "Microfluidic devices allow manipulation of reagents and fluids in a semi-automated fashion, ideal for performing multiple measurements or conditioning various reagents. Here, an enzyme assay has been performed in a multilayer poly(methyl methacrylate)-based microfluidic device, where the layers are fluidically connected via a nanocapillary array membrane serving as an effective injector and valve. As a model system, β-glucuronidase from Escherichia coli and fluorescein di(β-D-glucuronide) are used for the assay; offline mixing and online incubation of substrate and enzyme allow determination of the initial hydrolysis rates of the substrate under catalysis by β-glucuronidase. The Michaelis constant Km was determined to be ∼ 4.0 μM for the enzyme of 83 units/mL at ambient temperature. The 50% inhibitory concentration IC50 of D-saccharic acid-1,4-Iactone to 167 units/mL was estimated to be 3.0 μtM. These results demonstrate added functionality for a poly(methyl methacrylate)-based nanocapillary array membrane-containing microfluidic device for following enzyme reaction kinetics.",

keywords = "Enzyme kinetics, Microcontact printing, Microfluidics, Nanocapillary array membranes, Nanofluidics, Poly(methyl methacrylate)",

author = "Maojun Gong and Kim, {Bo Young} and Flachsbart, {Bruce R.} and Shannon, {Mark A.} and Bohn, {Paul W.} and Sweedler, {Jonathan V.}",

note = "Funding Information: Manuscript received June 1, 2007; accepted July 13, 2007. This work was supported in part by the National Science Foundation under Award DMI-0328162, in part by the Department of Energy under Grant DE FG02 07ER15851, and in part by the Strategic Environmental Research and Development Program under Award W9132T-05-2-0028. The associate editor coordinating the review of this paper and approving it for publication was Dr. Richard Fair.",

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An on-chip fluorogenic enzyme assay using a multilayer microchip interconnected with a nanocapillary array membrane

Abstract

Keywords

ASJC Scopus subject areas

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