Tracking Aggregation of Virus-Like Particles Using Dynamic Light Scattering

Resumen

Source: Moore, M. D., et al. Alternative In Vitro Methods for the Determination of Viral Capsid Structural Integrity. J. Vis. Exp. (2017)

The video demonstrates a protocol for monitoring the heat-induced aggregation of virus-like particles using dynamic light scattering (DLS). It begins with preparing a monodisperse suspension of purified particles. The sample is then loaded into a preheated DLS instrument. As the capsid proteins denature with heat, exposed hydrophobic regions cause particle aggregation. DLS detects changes in particle size over time, enabling real-time tracking of the aggregation process.

Protocolo

1. DLS for Detecting Virus Aggregation in Real Time

NOTE: The steps below may be specific to the software and instrument used, but can be adapted and applied to similar devices.

1. Create a new size SOP in the Dynamic Light Scattering (DLS) instrument software using: Material = Protein, Dispersant = Phosphate-buffered saline (PBS), Temperature = as desired, Equilibration time = 0 sec, Cell type = quartz cuvette, Measurement angle = 173° Backscatter, Measurement duration = automatic, Number of measurements = 50 (may be increased or decreased depending on aggregation rate), Delay between measurements = 0 sec, Data processing: Analysis mod....

Materiales

Lista de materiales utilizados en este artículo

Nombre	Empresa	Número de catálogo	Comentarios
Purified, assembled human norovirus GII.4 capsids
1x PBS (pH 7.2)	Life Technologies	20012-050
Zetasizer ZSP, or similar particle size analysis instrument	Malvern Instruments	N/A
Vortex	Fisher Scientifc	02-215-365
Quartz cuvette	Hellma	104-002-10-40
Small volume disposable cuvette (to reduce VLP use)	Malvern Instruments	ZEN0040