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An ELISA Lab-on-a-Chip (ELISA-LOC)

  • Avraham RasoolyEmail author
  • Hugh A. Bruck
  • Yordan Kostov
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 949)

Abstract

Laminated object manufacturing (LOM) technology using polymer sheets is an easy and affordable method for rapid prototyping of Lab-on-a-Chip (LOC) systems. It has recently been used to fabricate a miniature 96 sample ELISA lab-on-a-chip (ELISA-LOC) by integrating the washing step directly into an ELISA plate. LOM has been shown to be capable of creating complex 3D microfluidics through the assembly of a stack of polymer sheets with features generated by laser micromachining and by bonding the sheets together with adhesive. A six layer ELISA-LOC was fabricated with an acrylic (poly(methyl methacrylate) (PMMA)) core and five polycarbonate layers micromachined by a CO2 laser with simple microfluidic features including a miniature 96-well sample plate. Immunological assays can be carried out in several configurations (1 × 96 wells, 2 × 48 wells, or 4 × 24 wells). The system includes three main functional elements: (1) a reagent loading fluidics module, (2) an assay and detection wells plate, and (3) a reagent removal fluidics module. The ELISA-LOC system combines several biosensing elements: (1) carbon nanotube (CNT) technology to enhance primary antibody immobilization, (2) sensitive ECL (electrochemiluminescence) detection, and (3) a charge-coupled device (CCD) detector for measuring the light signal generated by ECL. Using a sandwich ELISA assay, the system detected Staphylococcal enterotoxin B (SEB) at concentrations as low as 0.1 ng/ml, a detection level similar to that reported for conventional ELISA. ELISA-LOC can be operated by a syringe and does not require power for operation. This simple point-of-care (POC) system is useful for carrying out various immunological assays and other complex medical assays without the laboratory required for conventional ELISA, and therefore may be more useful for global healthcare delivery.

Key words

ELISA Lamination Charge-coupled device Micromachining Microfluidics Staphylococcal enterotoxins Enhanced chemiluminescence Carbon nanotubes Point-of-care-settings Food safety 

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Copyright information

© Springer Science+Business Media,LLC 2013

Authors and Affiliations

  • Avraham Rasooly
    • 1
    • 2
    Email author
  • Hugh A. Bruck
    • 3
  • Yordan Kostov
    • 4
    • 5
  1. 1.Division of Biology, Office of Science and EngineeringFDA Center for Devices and Radiological Health (CDRH)Silver SpringUSA
  2. 2.National Cancer InstituteRockvilleUSA
  3. 3.Department of Mechanical EngineeringUniversity of MarylandCollege ParkUSA
  4. 4.Steven Sun Division of Biology Office of Science and EngineeringFDA Center for Devices and Radiological Health (CDRH)Silver SpringUSA
  5. 5.University of Maryland Baltimore CountyBaltimore CountyUSA

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