Directed Evolution of a G-Quadruplex Peroxidase DNAzyme and Application in Proteomic DNAzyme-Aptamer Proximity Labeling.

DNAzymes have been limited in application by their low catalytic rates. Here, we evolved a new peroxidase DNAzyme mSBDZ-X-3 through a directed evolution method based on the capture of self-biotinylated DNA catalyzed by its intrinsic peroxidase activity. The mSBDX-X-3 DNAzyme has a parallel G-quadruplex structure and has more favorable catalytic properties than all previously reported peroxidase DNAzyme variants. We applied mSBDZ-X-3 in an aptamer-coupled proximity-based labeling proteomic assay to determine the proteins that bind to cell surface cancer biomarkers EpCAM and nucleolin. Confocal microscopy, western blot analysis, and LC-MS/MS showed that the hybrid DNAzyme aptamer-coupled proximity assay-labeled proteins associated with EpCAM and nucleolin within 6-12 min in fixed cancer cells. The labeled proteins were identified by mass spectrometry. This study provides a highly efficient peroxidase DNAzyme, a methodology for selection of such variants, and a method for its application in spatial proteomics using entirely nucleic acid-based tooling.

Microfluidic Technology for Nucleic Acid Aptamer Evolution and Application.

The intersection of microfluidics and aptamer technologies holds particular promise for rapid progress in a plethora of applications across biomedical science and other areas. Here, the influence of microfluidics on the field of aptamers, from traditional capillary electrophoresis approaches through innovative modern-day approaches using micromagnetic beads and emulsion droplets, is reviewed. Miniaturizing aptamer-based bioassays through microfluidics has the potential to transform diagnostics and embedded biosensing in the coming years.

Sensitivity enhancement in swept-source optical coherence tomography by parametric balanced detector and amplifier.

We proposed a sensitivity enhancement method of the interference-based signal detection approach and applied it on a swept-source optical coherence tomography (SS-OCT) system through all-fiber optical parametric amplifier (FOPA) and parametric balanced detector (BD). The parametric BD was realized by combining the signal and phase conjugated idler band that was newly-generated through FOPA, and specifically by superimposing these two bands at a photodetector. The sensitivity enhancement by FOPA and parametric BD in SS-OCT were demonstrated experimentally. The results show that SS-OCT with FOPA and SS-OCT with parametric BD can provide more than 9 dB and 12 dB sensitivity improvement, respectively, when compared with the conventional SS-OCT in a spectral bandwidth spanning over 76 nm. To further verify and elaborate their sensitivity enhancement, a bio-sample imaging experiment was conducted on loach eyes by conventional SS-OCT setup, SS-OCT with FOPA and parametric BD at different illumination power levels. All these results proved that using FOPA and parametric BD could improve the sensitivity significantly in SS-OCT systems.

Tri-band spectroscopic optical coherence tomography based on optical parametric amplification for lipid and vessel visualization.

A tri-band spectroscopic optical coherence tomography (SOCT) system has been implemented for visualization of lipid and blood vessel distribution. The tri-band swept source, which covers output spectrum in 1.3, 1.5, and 1.6 µm wavelength windows, is based on a dual-band Fourier domain mode-locked laser and a fiber optical parametric amplifier. This tri-band SOCT can further differentiate materials, e.g., lipid and artery, qualitatively by contrasting attenuation coefficients difference within any two of these bands. Furthermore, ex vivo imaging of both porcine artery with artificial lipid plaque phantom and mice with coronary artery disease were demonstrated to showcase the capability of our SOCT.