A Novel Transgenic Sf9 Cell Line for Quick and Easy Virus Quantification

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Biology

Background:

• The baculovirus expression system (BES) is widely used for protein production.
• Accurate and rapid virus quantification is crucial for optimizing BES applications.
• Existing methods for virus quantification can be time-consuming.

Purpose of the Study:

• To generate a transgenic Sf9 cell line for enhanced and rapid virus quantification.
• To develop a cell line that facilitates easy and efficient analysis in BES.
• To improve the speed of virus quantification without compromising accuracy.

Main Methods:

• Utilized the piggyBac system to create transgenic Sf9 cell lines.
• Employed hr3, 39K, and p10 promoters for enhanced green fluorescent protein (EGFP) expression.
• Screened 20 transgenic lines to select the optimal performer (Sf9-QE).
• Characterized Sf9-QE cells for diameter, proliferation rate, and stability.

Main Results:

• Selected Sf9-QE cell line exhibited rapid and intense EGFP expression upon viral infection.
• Sf9-QE cells showed a smaller average diameter (16 μm) and 1.6x higher proliferation rate than Sf9 cells.
• Virus quantification using Sf9-QE cells yielded comparable results to other methods.
• Achieved complete virus quantification in 5.3–6.0 days, significantly reducing time (4–6 days shorter).
• Sf9-QE cell properties remained stable for at least 100 passages.

Conclusions:

• The Sf9-QE cell line offers a significant advancement for rapid and convenient virus quantification in BES.
• This cell line enables accurate virus quantification via fluorescence photometry within approximately 6.0–6.3 days.
• The stability and enhanced proliferation of Sf9-QE cells make them a valuable tool for biotechnology research.