Rapid and Economic Baculovirus Titer Determination Using a Novel Transgenic Sf9-QE Cell Line.

A baculovirus expression system (BES) for the production of recombinant proteins requires rapid and easy virus titer determination. In this study, a novel direct titration method was developed using a novel Sf9-QE cell line to easily and economically determine virus titers in a short time. This direct titration method can determine virus titers by directly counting the initially infected cells. This method requires the rapid identification of the initial virus-infected cells. The genome of Sf9-QE cells, which fluoresce upon virus infection, was found to contain at least seven copy numbers of the enhanced green fluorescent protein (EGFP) transgene. This result suggests that Sf9-QE cells in the early stages of virus infection can be identified by the high expression of EGFP. It was also shown that for accurate virus titration using the direct titration method, Sf9-QE cells should be used within 3 d of subculturing. Additionally, counting fluorescent cells to establish virus infection should be performed within 15 to 30 h after virus infection for reliable virus titration. The direct titration method using Sf9-QE cells provides a rapid, reliable, and cost-effective alternative for determining baculovirus titers in BES research.