Comparison of DNAzyme activity for the development of an immobilized heme sensor

Abstract

Point-of-care systems require highly sensitive, quantitative and selective detection platforms for the real-time multiplexed monitoring of target analytes. To ensure facile development of a sensor, it is preferable for the detection assay to have minimal chemical complexity, contain no wash steps and provide a wide and easily adaptable detection range for multiple targets. Current studies involve label-free detection strategy for relevant clinical molecules such as heme using G-quadruplex based self-assembly. We have explored the measurement of binding and kinetic parameters of various G-quadruplex/heme complexes which are able to self-associate to form a DNAzyme with peroxidase mimicking capabilities and are critical to nucleic acid research. The detection strategy includes immobilizing the G-quadruplex sequences within a polymer matrix to provide a self-assembly based detection approach for heme that could be translated towards other clinically relevant targets.

DNAzyme-Based Biosensors: Immobilization Strategies, Applications, and Future Prospective

“…Specifically, Chen et al immobilized a peroxidase mimicking DNAzyme onto eggshell membranes for chemiluminescent detection of hydrogen peroxide . Other studies have relied on pre-purchased reagents such as aldehyde-terminated agarose microbeads and aldehyde-coated glass slides for DNAzyme-based detection of heme and Pb 2+ , respectively. , …”

“…46 Other studies have relied on pre-purchased reagents such as aldehyde-terminated agarose microbeads and aldehyde-coated glass slides for DNAzyme-based detection of heme and Pb 2+ , respectively. 47,48 Alternatively, surface functionalization with esters provides another means by which aminated DNAzymes can be immobilized. Most prominently, this pathway involves inducing carboxyl groups onto the surface and subsequent treatment with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS).…”