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Technical Brief: HyQ® SFX-Insect™

Production of rAcMNPV stock

1.

Plant Sf9 cells in shakers, Fernbachs or bioreactor at 5 x 105 cells/mL and grow for 3-5 days in HyQ SFX-Insect until the cell density reaches 3-6 x 106 cells/mL with 95% viability.
2. Remove the spent medium by aspiration from sedimented/concentrated cells and resuspend to original volume in the resident vessel.
3. Subculture the cells and plant 1-2 x 106 cells/mL into appropriate vessels using fresh HyQ SFX-Insect. Incubate the cells in incubator for 1 hour for physiological and physio-chemical equilibrium. Perform cell count which should be about 95-98% viable. Infect the cells with with MOI = 0.5 of rAcMNPV.
4. Incubate for 48 hours and harvest sample for titration by PFU. (Budded Virus)
5. Concentrate the virus, plaque titrate and check for expression vs standard control. Stabilize the virus with glutamates or other suitable stabilizers and quick freeze using dry ice-ETOH method. Store the stock virus at -80°C or liquid N2.

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