Glucose oxidase-mediated polymerization as a platform for dual-mode signal amplification and biodetection

Brad J. Berron, Leah M. Johnson, Xiao Ba, Joshua D. McCall, Nicholas J. Alvey, Kristi S. Anseth, Christopher N. Bowman

Research output: Contribution to journalArticlepeer-review

46 Scopus citations


We report the first use of a polymerization-based ELISA substrate solution employing enzymatically mediated radical polymerization as a dual-mode amplification strategy. Enzymes are selectively coupled to surfaces to generate radicals that subsequently lead to polymerization-based amplification (PBA) and biodetection. Sensitivity and amplification of the polymerization-based detection system were optimized in a microwell strip format using a biotinylated microwell surface with a glucose oxidase (GOx)-avidin conjugate. The immobilized GOx is used to initiate polymerization, enabling the detection of the biorecognition event visually or through the use of a plate reader. Assay response is compared to that of an enzymatic substrate utilizing nitroblue tetrazolium in a simplified assay using biotinylated wells. The polymerization substrate exhibits equivalent sensitivity (2μg/mL of GOx-avidin) and over three times greater signal amplification than this traditional enzymatic substrate since each radical that is enzymatically generated leads to a large number of polymerization events. Enzyme-mediated polymerization proceeds in an ambient atmosphere without the need for external energy sources, which is an improvement upon previous PBA platforms. Substrate formulations are highly sensitive to both glucose and iron concentrations at the lowest enzyme concentrations. Increases in amplification time correspond to higher assay sensitivities with no increase in non-specific signal. Finally, the polymerization substrate generated a signal to noise ratio of 14 at the detection limit (156ng/mL) in an assay of transforming growth factor-beta.

Original languageEnglish
Pages (from-to)1521-1528
Number of pages8
JournalBiotechnology and Bioengineering
Issue number7
StatePublished - Jul 2011


  • Biological detection
  • Dual-mode amplification
  • Enzymatic polymerization

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology


Dive into the research topics of 'Glucose oxidase-mediated polymerization as a platform for dual-mode signal amplification and biodetection'. Together they form a unique fingerprint.

Cite this