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Related Experiment Video

Updated: Jun 22, 2025

The MultiBac Protein Complex Production Platform at the EMBL

The MultiBac Protein Complex Production Platform at the EMBL

Published on: July 11, 2013

Functional Baculovirus Particle Quantification via Plaque Assay.

Pranav R H Joshi¹, Alina Venereo-Sanchez²

¹Viral Vectors and Vaccines Bioprocessing Group, Department of Bioengineering, McGill University, Montreal, QC, Canada.

Methods in Molecular Biology (Clifton, N.J.)

|July 1, 2024

Summary

This summary is machine-generated.

This study details a plaque assay method for quantifying infectious baculovirus, crucial for determining viral doses for cell infections. The method accurately measures plaque-forming units (PFU) for recombinant baculovirus expression vectors.

Keywords:

Baculovirus Insect cell culture Plaque assay Plaque forming units Recombinant baculovirus expression vector rBEV

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Last Updated: Jun 22, 2025

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Area of Science:

Virology
Molecular Biology
Biotechnology

Background:

Accurate quantification of infectious baculovirus is essential for reproducible experiments.
The plaque assay is a standard method for determining viral titer as plaque-forming units (PFU).
Determining the multiplicity of infection (MOI) requires precise viral quantification.

Purpose of the Study:

To describe a plaque assay method for quantifying infectious baculovirus.
To enable the determination of viral stock concentration for cell culture applications.
To validate the method for quantifying recombinant baculovirus expression vectors (rBEVs).

Main Methods:

Serial dilutions of baculovirus stock were applied to Sf9 cell monolayers.
A 5% Agarose overlay was added post-infection.
Infection halos were visualized using neutral red solution six days post-infection.

Main Results:

The plaque assay method allowed for the quantification of infectious baculovirus as plaque-forming units (PFU).
The method enabled the determination of the precise amount of virus needed for a specific multiplicity of infection (MOI).
Reproducible quantification of PFU was achieved for recombinant baculovirus expression vectors (rBEVs).

Conclusions:

The described plaque assay is a reliable method for quantifying infectious baculovirus.
This technique is suitable for determining viral titers of recombinant baculovirus expression vectors.
Accurate PFU quantification is critical for successful baculovirus-mediated gene expression studies.