FRET Detection of Proteins Using Fluorescently Doped Electrospun Nanofibers and Pattern Recognition

Abstract

This paper reports the fabrication of solid-state nanofiber sensor arrays and their use for detection of multiple proteins using principal component analysis (PCA). Four cationic and anionic fluorescently embedded nanofibers are generated by an electrospinning method, yielding unique patterns of fluorescence change upon interaction with protein samples. Five metal and nonmetal containing proteins, i.e., hemoglobin, myoglobin, cytochrome c, BSA, and avidin, have been investigated; and the results show that distinct fluorescent patterns can be formed upon the addition of protein samples to the array of solid nanofiber substrates, allowing their unambiguous identification. The nanofiber films are highly repeatable with a batch-to-batch variation of approximately 5% and demonstrated outstanding reusability with less than a 15% loss of fluorescence intensity signal after 5 regenerations of test cycles. For a more practical visualization, a cluster map was generated using PCA of the change-in-fluorescence (ΔI) composite patterns, demonstrating the potential of the method for diagnostic applications.

Laser Systems and Networks with Organic Nanowires and Nanofibers

“…For instance, strategies for achieving spectral tunability in light‐emitting NFs include the use of calibrated light‐emitting blends with energy donors and acceptors that present stimulated emission and Förster resonance energy transfer . Methods for exploiting biocompatibility of organic light‐emitting NWs and NFs, as well as for deploying their extraordinary sensitivity for surrounding microenviromental conditions, include programming them to detect proteins, metal cations, or explosives . In this framework, microfiber lasers can be applied with success for refractive index sensing, based on the shift of lasing modes.…”

Light‐Emitting Electrospun Nanofibers for Nanophotonics and Optoelectronics

“…Other examples of optical sensors include nanofibers made by polyfluorene‐derivatives blended with PMMA, which display a high sensitivity toward plasmid DNA,56b and nanofibers made by a diphenylacetylene polymer, sensitive to explosive nitroaromatic compounds 120, 121. Optical sensors based on fluorescent nanofibers have also been developed for protein detection 122, 123. Wang et al124 have recently demonstrated ultrasensitive detection of explosives vapors, explosives residues on handprint and buried explosives, by means of emission quenching in a sensing film composed of electrospun nanofibers.…”

Materials for FRET Analysis: Beyond Traditional Dye–Dye Combinations

“…Inherently fluorescent cationic dendritic structures comprising primarily phenylene-ethynylene have been used as donors for the detection of peptide nucleic acid (PNA)/DNA hybridization, where the neutral PNA probe was labeled with a fluorescein acceptor [127]. Davis et al used inherently fluorescent cationic and anionic diphenylacetylene dendrimers doped into cellulose acetate to generate solid-state electrospun nanofiber sensor arrays for protein detection [128]. These nanofiber films selectively interacted with proteins (including metal-and nonmetal-containing proteins), which resulted in fluorescent quenching in distinct patterns, due to varying interactions with the different protein structures; this allows specific protein identification even in complex mixtures.…”