Generation of High-Titer Baculovirus in Insect Cells

Begin with a suspension culture of insect cells.

Add a baculovirus culture and incubate under agitation for uniform spread.

During incubation, the virus enters the cell, releases its nucleocapsid, and transports the viral DNA to the nucleus.

In the nucleus, the viral genome replicates to amplify viral DNA and undergoes transcription to produce mRNA.

The mRNA exits to the cytoplasm, directs structural protein synthesis, and the proteins enter the nucleus to assemble nucleocapsids around the newly replicated genomes.

The nucleocapsids exit the nucleus, and new virions bud off. The released virions infect neighboring cells, amplifying viral production.

As the infection progresses, increased intracellular viral protein levels cause cell enlargement, nuclear swelling, and lysis, indicating high-titer viral production.

Next, transfer the baculovirus-containing supernatant to fresh cells and incubate to amplify viral production.

Transfer the culture into a tube, centrifuge to separate the cells and debris, then collect the viral supernatant.

The high-titer baculovirus is ready for purification.

To set up the suspension culture, seed 50 milliliters of Sf9 cells in insect culture medium into a 250-mi