Systematic Study of Fluorescein-Functionalized Macrophotoinitiators for Colorimetric Bioassays

We report a systematic investigation of a set of macrophotoinitiators for use in polymerization-based signal amplification. To test the dependence of photopolymerization responses on the number of photoinitiators localized per molecular recognition event, we gradually increased the number of photoinitiator molecules coupled to a scaffold macromolecule. Macrophotoinitiators constructed with an average of 7 to 168 photoinitiators per polymer with the goals of quantifying the relationship between the number of initiators per binding event and the degree of amplified colorimetric readout. To evaluate the capacity of the macrophotoinitiators to detect molecular recognition, neutravidin was coupled to these molecules to recognize biotin-labeled DNA immobilized on biochip test surfaces. Fluorescein macroinitiators are found to be useful in detecting molecular recognition above a threshold of initiators per polymer. Above this threshold, increasing the number of initiators per macroinitiator resulted in increased signal strength.