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Production and Purification of Baculovirus for Gene Therapy Application
Published on: April 9, 2018
Estimation of baculovirus titer based on viable cell size
Vasantharajan Janakiraman1, William F Forrest, Somasekar Seshagiri
1Department of Molecular Biology, Genentech Inc., 1 DNA way, South San Francisco, California 94080, USA.
Nature Protocols
|April 5, 2007
Summary
This study introduces a rapid method to determine baculovirus titers by measuring insect cell size changes post-infection. This technique offers a faster alternative to traditional methods, completing in just 28 hours.
Area of Science:
- Virology
- Cell Biology
- Biotechnology
Background:
- Traditional baculovirus titration methods like plaque assays and qPCR are time-consuming and labor-intensive.
- Accurate baculovirus titer determination is crucial for efficient viral vector applications and research.
Purpose of the Study:
- To present a simple, rapid, and efficient protocol for determining baculovirus titers.
- To establish a new method based on measurable changes in insect cell size following infection.
Main Methods:
- Utilized the Vi-CELL instrument to measure changes in insect cell diameter after baculovirus infection across various dilutions.
- Employed the method of moments, a statistical approach, to compute virus titers from cell diameter data.
Main Results:
- The novel protocol allows for baculovirus titer determination within approximately 28 hours.
- Demonstrated a correlation between increasing viable insect cell size and baculovirus infection levels.
Conclusions:
- The presented cell size-based method provides a significantly faster and less labor-intensive alternative for baculovirus titration.
- This protocol can streamline research and applications involving baculoviruses, improving efficiency.
