Increase of reaction rate and sensitivity of low-abundance enzyme assay using micro/nanofluidic preconcentration chip

Jeong Hoon Lee, Rafael Song, Steven R. Tannenbaum, Jongyoon Han

    Research output: Contribution to journalArticle

    Abstract

    We report a novel method of increasing both the reaction rate and the sensitivity of low-abundance enzyme assay using a micro/nanofluidic preconcentration chip. The disposable preconcentration device made out of PDMS with a surface-patterned ion-selective membrane increases local enzyme/substrate concentrations for rapid monitoring of enzyme activity. As a model system, we used trypsin as the enzyme and BODIPY FL casein as the fluorogenic substrate. We demonstrated that the reaction rate of trypsin-BODIPY FL was significantly enhanced by increasing the local concentrations of both trypsin and BODIPY FL casein in the preconcentration chip. The reaction time required to turn over substrates at 1 ng/mL was only ∼10 min compared to ∼1 h without preconcentration, which demonstrates a significantly higher reaction rate through the increase of the concentrations of both the enzyme and substrate. Furthermore, trypsin activity can be measured down to a concentration level of 10 pg/mL, which is a ∼100 fold enhancement in sensitivity compared to the result without the preconcentration step. This micro/nanofluidic preconcentrator chip could be used as a generic micro reaction platform to study any enzyme-substrate systems, or other biochemical reaction systems in low concentration ranges.

    Original languageEnglish (US)
    Pages (from-to)3198-3204
    Number of pages7
    JournalAnalytical Chemistry
    Volume80
    Issue number9
    DOIs
    StatePublished - May 1 2008

    Fingerprint

    Nanofluidics
    Reaction rates
    Assays
    Trypsin
    Enzymes
    Substrates
    Caseins
    Ion selective membranes
    Enzyme activity
    Fluorescent Dyes
    Monitoring
    4,4-difluoro-4-bora-3a,4a-diaza-s-indacene

    ASJC Scopus subject areas

    • Analytical Chemistry

    Cite this

    Increase of reaction rate and sensitivity of low-abundance enzyme assay using micro/nanofluidic preconcentration chip. / Lee, Jeong Hoon; Song, Rafael; Tannenbaum, Steven R.; Han, Jongyoon.

    In: Analytical Chemistry, Vol. 80, No. 9, 01.05.2008, p. 3198-3204.

    Research output: Contribution to journalArticle

    Lee, Jeong Hoon ; Song, Rafael ; Tannenbaum, Steven R. ; Han, Jongyoon. / Increase of reaction rate and sensitivity of low-abundance enzyme assay using micro/nanofluidic preconcentration chip. In: Analytical Chemistry. 2008 ; Vol. 80, No. 9. pp. 3198-3204.
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