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Updated: Jul 14, 2026
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Detecting and Characterizing Protein Self-Assembly In Vivo by Flow Cytometry
Published on: July 17, 2019
Prototype protein assembly as scaffold for time-resolved fluoroimmuno assays.
Hannah N Barnhill1, Stéphanie Claudel-Gillet, Raymond Ziessel
1Department of Chemistry and Biochemistry and Nanocenter, University of South Carolina, 631 Sumter St., Columbia, South Carolina, USA.
Journal of the American Chemical Society
|June 5, 2007
Summary
Turnip yellow mosaic virus (TYMV) serves as a bionanoparticle scaffold for developing sensors. This study successfully dual-modified TYMV with luminescent terbium complexes and biotin for enhanced time-resolved fluoroimmuno assays (TRFIA).
Area of Science:
- Nanotechnology
- Bioconjugation
- Biophysics
Background:
- Turnip yellow mosaic virus (TYMV) is an inexpensive, abundant icosahedral plant virus.
- Bionanoparticles offer versatile platforms for developing advanced diagnostic tools.
- Orthogonal chemical strategies enable precise modification of biomolecular surfaces.
Purpose of the Study:
- To functionalize TYMV with luminescent terbium complexes and biotin groups.
- To investigate the spectroscopic properties and FRET efficiency of dual-modified TYMV.
- To evaluate TYMV as a scaffold for time-resolved fluoroimmuno assays (TRFIA).
Main Methods:
- Dual-modification of TYMV using orthogonal chemical reactions for Tb complex and biotin anchoring.
- Characterization of bioconjugation using Mass Spectrometry (MS) and Western blot.
- Spectroscopic analysis including steady-state and time-resolved luminescence.
- Fluorescence Resonance Energy Transfer (FRET) experiments with avidin-Alexa488.
Main Results:
- Successful dual-modification of TYMV without perturbing Tb complex spectroscopic properties.
- Demonstrated FRET between Tb complexes (donor) and Alexa488 (acceptor) with 66% energy transfer.
- Estimated average donor-acceptor distance of 36 Å.
- Confirmed unimpeded ligand-receptor binding (biotin-avidin) on the TYMV surface.
Conclusions:
- TYMV is a suitable prototype bionanoparticle for sensor development.
- Dual-modified TYMV enables efficient TRFIA applications.
- The study highlights the potential of nanoscale biological assemblies for functional sensor design.