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Creating advanced multifunctional biosensors with surface enzymatic transformations.

Hye Jin Lee1, Alastair W Wark, Robert M Corn

  • 1Department of Chemistry, University of California-Irvine, Irvine, California 92697, USA.

Langmuir : the ACS Journal of Surfaces and Colloids
|May 31, 2006
PubMed
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We developed novel multiplexed biosensors by combining surface enzyme chemistry and bioaffinity interactions on microarrays. This approach enables ultrasensitive DNA detection and on-chip RNA replication for advanced diagnostics.

Area of Science:

  • Biotechnology
  • Biosensor Development
  • Surface Chemistry

Background:

  • Multiplexed biosensors require enhanced selectivity and sensitivity.
  • Surface enzyme chemistry and bioaffinity interactions are key for biosensor performance.
  • Real-time detection of enzymatic transformations is crucial for biosensor applications.

Purpose of the Study:

  • To develop novel multiplexed biosensors by coupling surface enzyme chemistry and bioaffinity interactions.
  • To demonstrate ultrasensitive detection of genomic DNA and on-chip RNA replication.
  • To establish a theoretical framework for quantitating surface enzyme reactivity.

Main Methods:

  • Utilized surface plasmon resonance imaging (SPRI) and surface plasmon fluorescence spectroscopy (SPFS) for real-time detection.

Related Experiment Videos

  • Employed ribonuclease H (RNase H) for enzymatic amplification and DNA detection.
  • Applied RNA-DNA ligation and T7 RNA polymerase for microarray regeneration and replication.

    • Main Results:

    • Achieved ultrasensitive direct detection of genomic DNA at 1 fM without labeling or PCR amplification.
    • Demonstrated renewable RNA microarrays created from single-stranded DNA microarrays.
    • Successfully applied a theoretical model to quantify surface enzyme reactivity using SPRI and SPFS.

    Conclusions:

    • Coupling surface enzyme chemistry and bioaffinity interactions on biopolymer microarrays creates highly selective and sensitive multiplexed biosensors.
    • The developed methods enable ultrasensitive DNA detection and efficient on-chip RNA manipulation.
    • The theoretical framework provides accurate quantitation of surface enzyme kinetics for biosensor optimization.