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A rapid method for estimation of baculovirus titer based on viable cell size
Vasantharajan Janakiraman1, William F Forrest, Bernard Chow
1Department of Molecular Biology, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
Journal of Virological Methods
|October 11, 2005
Summary
A new method rapidly determines baculovirus titers by measuring cell size changes after infection, offering a faster alternative to traditional plaque assays for optimizing recombinant protein production.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Baculovirus expression systems are vital for recombinant protein production.
- Accurate virus titration is crucial for optimizing protein expression via multiplicity of infection (MOI).
- Existing methods like plaque assays are often time-consuming and labor-intensive.
Purpose of the Study:
- To develop a rapid and simple method for baculovirus titration.
- To optimize conditions for efficient recombinant protein expression.
- To establish a viable cell-diameter based method for virus titer estimation.
Main Methods:
- Utilized the Vi-CELL instrument to measure viable cell diameter post-baculovirus infection across various virus dilutions.
- Employed statistical modeling techniques to analyze cell diameter data.
- Compared results with traditional plaque assay titration methods.
Main Results:
- Demonstrated that viable cell diameter increase correlates with baculovirus infection.
- The cell-diameter based method provides reliable virus titer estimations.
- Results were comparable to those obtained from conventional plaque assays.
Conclusions:
- Viable cell diameter measurement offers a simple, rapid, and reliable alternative for baculovirus titration.
- This method can streamline the optimization of recombinant protein expression.
- Facilitates efficient MOI determination for enhanced protein yields.