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Design of a single-step immunoassay principle based on the combination of an enzyme-labeled antibody release coating and a hydrogel copolymerized with a fluorescent enzyme substrate in a microfluidic capillary device

Lab Chip. 2013 Nov 21;13(22):4304-7. doi: 10.1039/c3lc50775k.

Abstract

A combination of an enzyme-labeled antibody release coating and a novel fluorescent enzyme substrate-copolymerized hydrogel in a microchannel for a single-step, no-wash microfluidic immunoassay is demonstrated. This hydrogel discriminates the free enzyme-conjugated antibody from an antigen-enzyme-conjugated antibody immunocomplex based on the difference in molecular size. A selective and sensitive immunoassay, with 10-1000 ng mL(-1) linear range, is reported.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / chemistry
  • Alkaline Phosphatase / metabolism
  • Antibodies / chemistry
  • Antibodies / immunology*
  • Antigens / chemistry
  • Antigens / immunology
  • Antigens / metabolism
  • Coumarins / chemistry
  • Enzymes / chemistry*
  • Enzymes / metabolism
  • Enzymes, Immobilized / chemistry
  • Enzymes, Immobilized / metabolism
  • Fluorescent Dyes / chemistry*
  • Humans
  • Hydrogel, Polyethylene Glycol Dimethacrylate / chemistry*
  • Immunoassay / instrumentation
  • Immunoassay / methods*
  • Microfluidic Analytical Techniques* / instrumentation
  • Polymerization
  • Substrate Specificity

Substances

  • Antibodies
  • Antigens
  • Coumarins
  • Enzymes
  • Enzymes, Immobilized
  • Fluorescent Dyes
  • Hydrogel, Polyethylene Glycol Dimethacrylate
  • Alkaline Phosphatase