4. FBS in the medium neutralizes/inhibits the trypsin’s activity. It
is crucial to remove the remaining trypsin from the cells.
Bioreactor preparation
5. The tips of electrodes (e.g., oxygen electrode) may have a
concave form which can lead to the entrapment of air/oxygen
bubbles. Make sure, that the tip of the electrode is installed
below the stirred bubbles in the bioreactor.
All sampling tubes are to be connected with Luer lock
adapters, including all bottles (acid, base, medium, inoculum,
etc.).
6. (a) Do not autoclave the amplifier of the dielectric
spectroscopy.
(b) Open tubing clamps during autoclave so that the hot steam
reaches all surfaces.
(c) Add 10–20 mL PBS into the 1 L bioreactor to prevent a
drying-out of the pH electrode.
7. Do not autoclave the medium, or any of the medium supplements, as essential components will denature irreversibly due to
heat and pressure exposure.
8. Fill up the medium under the clean bench to avoid the risk of
contamination.
9. Set the maximum heating temperature in the thermal loop to
39–40
C to avoid a denaturation of medium components
(e.g., proteins) by overheating when warming up the medium
to a temperature of 37
C.
10. Prepare a 1 M NaOH solution, and sterilize it with a 0.2 μm
filter. Use a sterile filter in the lid, and attach silicon tubes. Set
up the silicon tube end with a Luer lock adapter to connect
these to the bioreactor.
Nanoplex production in the STR
11. Start the cell infection after the cells have completed their
attachment to the microcarrier.
Downstream processing
Initial sample clarification
12. An immediate clarification of the complete harvest is highly
recommended. A storage of the harvest can lead to an additional cell disruption, releasing enzymes that modify the target
an increase the product contact time to these enzymes. This can
potentially affect virus infectivity and stability.
13. Membrane-based filtration (depth and tangential flow filtration) is the method of choice for the clarification of larger
volumes. However, for small scale samples, centrifugation
might be considered.
Viral Nanoplex Vaccine Production
239
is crucial to remove the remaining trypsin from the cells.
Bioreactor preparation
5. The tips of electrodes (e.g., oxygen electrode) may have a
concave form which can lead to the entrapment of air/oxygen
bubbles. Make sure, that the tip of the electrode is installed
below the stirred bubbles in the bioreactor.
All sampling tubes are to be connected with Luer lock
adapters, including all bottles (acid, base, medium, inoculum,
etc.).
6. (a) Do not autoclave the amplifier of the dielectric
spectroscopy.
(b) Open tubing clamps during autoclave so that the hot steam
reaches all surfaces.
(c) Add 10–20 mL PBS into the 1 L bioreactor to prevent a
drying-out of the pH electrode.
7. Do not autoclave the medium, or any of the medium supplements, as essential components will denature irreversibly due to
heat and pressure exposure.
8. Fill up the medium under the clean bench to avoid the risk of
contamination.
9. Set the maximum heating temperature in the thermal loop to
39–40
C to avoid a denaturation of medium components
(e.g., proteins) by overheating when warming up the medium
to a temperature of 37
C.
10. Prepare a 1 M NaOH solution, and sterilize it with a 0.2 μm
filter. Use a sterile filter in the lid, and attach silicon tubes. Set
up the silicon tube end with a Luer lock adapter to connect
these to the bioreactor.
Nanoplex production in the STR
11. Start the cell infection after the cells have completed their
attachment to the microcarrier.
Downstream processing
Initial sample clarification
12. An immediate clarification of the complete harvest is highly
recommended. A storage of the harvest can lead to an additional cell disruption, releasing enzymes that modify the target
an increase the product contact time to these enzymes. This can
potentially affect virus infectivity and stability.
13. Membrane-based filtration (depth and tangential flow filtration) is the method of choice for the clarification of larger
volumes. However, for small scale samples, centrifugation
might be considered.
Viral Nanoplex Vaccine Production
239