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

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.

Comparison of eosin and fluorescein conjugates for the photoinitiation of cell-compatible polymer coatings

“…Importantly, this system has been used to initiate the bulk gelation of cell laden tissue engineering scaffolds with reported high cell viabilities [ 9 ]. Eosin Y has also been functionalized to surfaces to form hydrogel sites of protein recognition on microarray[ 10 – 12 ] and cellular substrates [ 1 ], and has shown remarkable efficiency and sensitivity in response to ultralow analyte densities [ 10 , 11 , 13 – 15 ].…”

Chitosan/Ce(IV) redox polymerization-based amplification for detection of DNA point mutation

“…Taking advantage of monomer replicating concept, polymerization‐based amplification (PBA) has recently emerged as a promising nonenzymatic signal amplification technique for use in molecular diagnostic devices . Amplification is achieved through the initiation of radical polymerization coupled to molecular recognition at a surface.…”

“…An initiator‐functionalized binding partner for the analyte molecule of interest and aqueous monomers that react to form a crosslinked hydrogel are the key chemical components of PBA. In recent years, various chemistries have been implemented, including atom transfer radical polymerization (ATRP), photo‐polymerization and reversible addition‐fragmentation chain transfer polymerization (RAFT), and have shown their promise as a point‐of‐care technology.…”

The Ever Changing Faces of Polymer Science: The 32nd Australasian Polymer Symposium

“…Professor Christopher Bowman from the University of Colorado had various research collaborations with different Australian researchers over many years. He frequently visited Australia and also attended many Australian Polymer Symposia, [17] often together with his research group. He became a true friend of many Australian scientists.…”