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High-throughput microarray-based enzyme-linked immunosorbent assay (ELISA).

L G Mendoza1, P McQuary, A Mongan

  • 1Genometrix, The Woodlands, TX, USA. lmendoza@genometrix.com

Biotechniques
|October 19, 1999
PubMed
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A novel biochip enables high-throughput, multiplexed enzyme-linked immunosorbent assays (ELISAs) using a 96-well format. This technology facilitates rapid, low-cost detection of protein antigens for diverse diagnostic applications.

Area of Science:

  • Biotechnology
  • Assay Development
  • Microarray Technology

Background:

  • Traditional enzyme-linked immunosorbent assays (ELISAs) can be time-consuming and limited in multiplexing capabilities.
  • High-throughput screening (HTS) is crucial for accelerating diagnostics and drug discovery.
  • Existing microarray formats may lack compatibility with automated robotic systems.

Purpose of the Study:

  • To describe a new generation biochip designed for high-throughput (HT), multiplexed ELISAs.
  • To demonstrate the feasibility of specific multiplex detection of protein antigens on a glass substrate.
  • To present a low-cost, highly parallel assay format compatible with automated systems.

Main Methods:

  • Development of a biochip with 96 wells, each containing 36-element arrays (144 elements/well) of 8 antigens and a marker protein.

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  • Utilized a custom capillary-based print head on an X-Y-Z robot for array fabrication (capacity >20,000 arrays/day).
  • Employed a scanning charge-coupled device (CCD) detector for quantitative imaging of 96 arrays in 30 seconds.

    • Main Results:

    • Demonstrated successful individual and collective detection of arrayed antigens using standard ELISA techniques.
    • Confirmed the feasibility of specific multiplex detection of protein antigens on a glass substrate.
    • The 96-well open microarray architecture is compatible with automated robotic systems.

    Conclusions:

    • The new biochip technology supports a low-cost, highly parallel assay format for multiplexed ELISAs.
    • This HTS format has potential applications in cellular protein expression profiling.
    • Future uses include multiplexed assays for infectious agents and cancer diagnostics.