Characterization of the Assaying Methods in Polymerization-Based Amplification of Surface Biomarkers

Leah M. Johnson; Heather J. Avens; Ryan R. Hansen; Holly L. Sewell; Christopher N. Bowman

doi:10.1071/CH09095

RESEARCH FRONT

Previous Next Contents Vol 62(8)

Characterization of the Assaying Methods in Polymerization-Based Amplification of Surface Biomarkers

Leah M. Johnson ^B , Heather J. Avens ^A , Ryan R. Hansen ^A , Holly L. Sewell ^A and Christopher N. Bowman ^A ^C

+ Author Affiliations

- Author Affiliations

^A Department of Chemical and Biological Engineering, University of Colorado, ECCH 111, CB 424, Boulder, CO 80309, USA.

^B Department of Chemistry and Biochemistry, University of Colorado, UCB 215, Boulder, CO 80309, USA.

^C Corresponding author. Email: christopher.bowman@colorado.edu

Australian Journal of Chemistry 62(8) 877-884 https://doi.org/10.1071/CH09095
Submitted: 15 February 2009 Accepted: 2 May 2009 Published: 13 August 2009

Abstract

Polymerization-based amplification (PBA), which combines bio-recognition events with polymerization reactions on surfaces, provides visual, sensitive, and cost-effective detection of biological interactions, particularly at extremely low levels of the targeted biological moiety. This study characterizes assay parameters that enhance the utility of PBA to detect nucleic acid and protein biomarkers. Here, we successfully employ PBA on surfaces that contain uniform, high density, immobilized capture molecules, including three-dimensional nitrocellulose-coated substrates. Optimized assay and polymerization conditions are used to characterize the dynamic polymer film heights on glass substrates that result from solutions of KRAS proto-oncogene biomarker targets at concentrations between 5 nM and 500 pM. Differing aqueous monomer formulations are utilized to produce 20 nm films at the 500 pM DNA detection limit.

Acknowledgements

This work has been supported by the State of Colorado and the University of Colorado Technology Transfer Office and by the National Institutes of Health 1R21 CA 127884. This work has also been supported by a National Science Foundation Graduate Research Fellowship to H.J.A. L.M.J., H.J.A., and R.R.H. would like to acknowledge the Graduate Assistantship in Areas of National Need Fellowship from the USA Department of Education. R.R.H. would also like to acknowledge the Teets Family Endowed Doctoral Fellowship in Nanotechnology for financial support of this work.

References

Characterization of the Assaying Methods in Polymerization-Based Amplification of Surface Biomarkers

Abstract

Acknowledgements

Subscriber Login