Signal amplification is a critical component of any sensitive analyte detection assay. Chain polymerization reactions offer a chemical approach to signal amplification, as a single initiation event can incite formation of a long polymer chain. We have developed a new analyte detection assay based on aqueous atom transfer radical polymerization (ATRP). This approach provides real-time readouts of polymerization progress by applying fluorogenic monomers – monomers that are dark in their monomer form but fluoresce under UV light when incorporated into the polymer chain – to couple the presence of analyte to polymer fluorescence in solution. Though encouraging, the practicality of the originally reported fluorogenic ATRP approach was limited by long reaction times for detectable fluorescence generation. To increase the polymerization kinetics of our fluorogenic ATRP reaction platform, we have conducted a systematic optimization study to probe the effect of several fluorogenic ATRP reaction variables, including catalyst/ligand loading, halide salt, reducing agent concentration, solvent and added surfactant on the kinetics of fluorescence generation and polymer composition. Combining the effects of optimized individual reaction components affords a significant increase in both the rate and magnitude of fluorescence generation more suitable for biodetection applications.

Fluorogenic atom transfer radical polymerization (ATRP) for analyte detection

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