Expression - Baculovirus Titering Kit—Determine the Amount of Infectious Baculovirus

Staining of cells with a fluorescently labeled anti-gp64 antibody allows for identification of infected insect cells. By inoculating cultures with a series of log dilutions of virus, and staining the cultures 18–20 hours post inoculation, the ratio of infected to uninfected insect cells can be determined by flow cytometry. Statistical analysis of the percentage of infected cells in the virus dilution series enables accurate infectious titer determination.

Alternative assays that also claim results in under 24 hours (such as particle counting instruments and qPCR) are indeed fast but do not deliver true infectious titer values. In contrast to the plaque assay, the culturing conditions employed for the flow cytometric assay closely reflect actual culturing conditions utilized for expression cultures, resulting in a more relevant titer value in a fraction of the time. Provided access to a flow cytometer equipped to handle 96 well plates, the flow cytometric assay is easily adapted to high throughput processes.

• Reproducible and accurate titer results within 24 hours
• Measures infectious virus
• Suspension culture methodology simulates actual production conditions
• Works with any flow cytometer capable of reading samples from 96-well plates
• Flow cytometric data collection removes the operator error of counting plaques
• gp64-PE antibody can be used alone to monitor infection kinetics of baculovirus in insect cell culture

gp64-PE (R-PHYCOERYTHRIN (R-PE)-CONJUGATED MOUSE ANTI-BACULOVIRUS MONOCLONAL ANTIBODY)

• Clone: AcV1
• Antibody Composition: Mouse IgG2A
• Storage Buffer: Phosphate buffered saline (PBS) containing 0.05% BSA
• and 0.09% sodium azide.

Recombinant Baculovirus Control

• BestBac 2.0 co-transfected with empty pVL1393 produced in Sf9 cells