0000000000390661
AUTHOR
Janak Prasad
Plasmonic Nanosensors for the Determination of Drug Effectiveness on Membrane Receptors.
We demonstrate the potential of the NanoSPR (nanoscale surface plasmon resonance sensors) method as a simple and cheap tool for the quantitative study of membrane protein–protein interactions. We use NanoSPR to determine the effectiveness of two potential drug candidates that inhibit the protein complex formation between FtsA and ZipA at initial stages of bacterial division. As the NanoSPR method relies on individual gold nanorods as sensing elements, there is no need for fluorescent labels or organic cosolvents, and it provides intrinsically high statistics. NanoSPR could become a powerful tool in drug development, drug delivery, and membrane studies.
Single Unlabeled Protein Detection on Individual Plasmonic Nanoparticles
The ultimate detection limit in analytic chemistry and biology is the single molecule. Commonly, fluorescent dye labels or enzymatic amplification are employed. This requires additional labeling of the analyte, which modifies the species under investigation and therefore influences biological processes. Here, we utilize single gold nanoparticles to detect single unlabeled proteins with extremely high temporal resolution. This allows for monitoring the dynamic evolution of a single protein binding event on a millisecond time scale. The technique even resolves equilibrium coverage fluctuations, opening a window into Brownian dynamics of unlabeled macromolecules. Therefore, our method enables …
Multiplexed plasmon sensor for rapid label-free analyte detection.
Efficient and cost-effective multiplexed detection schemes for proteins in small liquid samples would bring drastic advances to fields like disease detection or water quality monitoring. We present a novel multiplexed sensor with randomly deposited aptamer functionalized gold nanorods. The spectral position of plasmon resonances of individual nanorods, monitored by dark-field spectroscopy, respond specifically to different proteins. We demonstrate nanomolar sensitivity, sensor recycling, and the potential to upscale to hundreds or thousands of targets.
Plasmonic Core–Satellite Assemblies as Highly Sensitive Refractive Index Sensors
Highly sensitive and spectrally tunable plasmonic nanostructures are of great demand for applications such as SERS and parallel biosensing. However, there is a lack of such nanostructures for the midvisible spectral regions as most available chemically stable nanostructures offer high sensitivity in the red to far red spectrum. In this work, we report the assembly of highly sensitive nanoparticle structures using a hydroxylamine mediated core–satellite assembly of 20 nm gold nanoparticle satellites onto 60 nm spherical gold cores. The average number of satellites allows tuning the plasmon resonance wavelength from 543 to 575 nm. The core–satellite nanostructures are stable in pH ranges from…
Plasmonic Nanosensors for Simultaneous Quantification of Multiple Protein–Protein Binding Affinities
Most of current techniques used for the quantification of protein-protein interactions require the analysis of one pair of binding partners at a time. Herein we present a label-free, simple, fast, and cost-effective route to characterize binding affinities between multiple macromolecular partners simultaneously, using optical dark-field spectroscopy and individual protein-functionalized gold nanorods as sensing elements. Our NanoSPR method could easily become a simple and standard tool in biological, biochemical, and medical laboratories.
Conformational dynamics of a single protein monitored for 24 hours at video rate
We use plasmon rulers to follow the conformational dynamics of a single protein for up to 24 h at a video rate. The plasmon ruler consists of two gold nanospheres connected by a single protein linker. In our experiment, we follow the dynamics of the molecular chaperone heat shock protein 90 (Hsp90), which is known to show “open” and “closed” conformations. Our measurements confirm the previously known conformational dynamics with transition times in the second to minute time scale and reveals new dynamics on the time scale of minutes to hours. Plasmon rulers thus extend the observation bandwidth 3–4 orders of magnitude with respect to single-molecule fluorescence resonance energy transfer a…