Nanoparticleâ€Templated Molecular Recognition Platforms for Detection of Biological Analytes

Molecular recognition of biological analytes with optical nanosensors provides both spatial and temporal biochemical information. A recently developed sensing platform exploits near-infrared fluorescent single-wall carbon nanotubes combined with electrostatically pinned heteropolymers to yield a synthetic molecular recognition technique that is maximally transparent through biological matter. This molecular recognition technique is known as corona phase molecular recognition (CoPhMoRe). In CoPhMoRe, the specificity of a folded polymer toward an analyte does not arise from a pre-existing polymer-analyte chemical affinity. Rather, specificity is conferred through conformational changes undergone by a polymer that is pinned to the surface of a nanoparticle in the presence of an analyte and the subsequent modifications in fluorescence readout of the nanoparticles. The protocols in this article describe a novel single-molecule microscopy tool (near-infrared fluorescence and total internal reflection fluorescence [nIRF TIRF] hybrid microscope) to visualize the CoPhMoRe recognition process, enabling a better understanding of synthetic molecular recognition. We describe this requisite microscope for simultaneous single-molecule visualization of optical molecular recognition and signal transduction. We elaborate on the general procedures for synthesizing and identifying single-walled carbon nanotube-based sensors that employ CoPhMoRe via two biologically relevant examples of single-molecule recognition for the hormone estradiol and the neurotransmitter dopamine. ; © 2016 John Wiley & Sons. Issue Online: 13 September 2016. Version of Record online: 13 September 2016. This work was supported by a Burroughs Wellcome Fund Career Award at the Scientific Interface (CASI), a Brain and Behavior Research foundation young investigator grant, and a Beckman Foundation Investigator Award. A.G.B. acknowledges a U.C. Berkeley Chancellor's Fellowship and an NSF Graduate Research Fellowship. G.S.D. acknowledges a Schlumberger ...