2.2 Downstream
Processing
2.2.1 Harvest of the Viral
Nanoplex-Containing
Supernatant, and Primary
Clarification
1. 1-L glass STR, working volume: 0.5 L (model number
Z611000110; Applikon, Biotechnology, Delft, Netherlands)—filled with microcarriers, cell culture medium, Vero
cells (grow on microcarrier) and viral nanoplexes in
supernatant.
2. SciPress Sensors (Parker Hannifin Manufacturing Ltd., Durham, England).
3. Balance (TE4101, Sartorius Stedim Biotech GmbH, Go ¨ttingen, Germany).
4. WinWedge software (Tal Technologies Inc., Philadelphia,
PA, USA).
5. Peristaltic pump (Sartorius Stedim Biotech GmbH, Go ¨ttingen,
Germany) with Tandem 1082 pump head, with a Masterflex
Pharmed #15 (Cole Parmer GmbH, Wertheim, Germany)
tube and transparent silicon tubes (VWR International, Radnor, PA, USA) with sterile Luer lock connections (Sarstedt,
Nu ¨mbrecht, Germany).
6. Opticap XL 1 Capsule with Polygard
® -CR 5.0 μm (Nominal)—sterile (Merck KGaA, Darmstadt, Germany).
7. 1 L PBS—Sterile (#L1825, Biochrom, Berlin, Germany).
8. Sodium hydroxide (Carl Roth GmbH, Karlsruhe, Germany).
9. Sterile bottles (0.5 L, Schott AG, Mainz, Germany).
10. Sterile workbench (ESCO Infinity Class II, Esco Lifesciences
GmbH, Friedberg, Germany).
2.2.2 Preparation
of Buffers and Membranes
1. PBS (#L1825, Biochrom, Berlin, Germany).
2. Deionized water (Milli-Q Academic, Q-Gard
® 1, Merck
KGaA, Darmstadt, Germany).
3. PEG 8000 (biotechnology grade, VWR International, Radnor,
PA, USA).
4. Sodium chloride (Sigma-Aldrich, Hamburg, Germany).
5. Sodium hydroxide 1 M NaOH (Carl Roth GmbH, Karlsruhe,
Germany).
6. 100-mL, 250-mL, and 1-L glass beakers (Carl Roth GmbH,
Karlsruhe, Germany).
7. 100-mL, 250-mL, and 500-mL measuring cylinders (Carl
Roth GmbH, Karlsruhe, Germany).
8. Analytical scale (Kern & Sohn GmbH, Balingen-Frommern,
Germany).
9. Magnetic stirring bar (Carl Roth GmbH, Karlsruhe,
Germany).
Viral Nanoplex Vaccine Production
227
Processing
2.2.1 Harvest of the Viral
Nanoplex-Containing
Supernatant, and Primary
Clarification
1. 1-L glass STR, working volume: 0.5 L (model number
Z611000110; Applikon, Biotechnology, Delft, Netherlands)—filled with microcarriers, cell culture medium, Vero
cells (grow on microcarrier) and viral nanoplexes in
supernatant.
2. SciPress Sensors (Parker Hannifin Manufacturing Ltd., Durham, England).
3. Balance (TE4101, Sartorius Stedim Biotech GmbH, Go ¨ttingen, Germany).
4. WinWedge software (Tal Technologies Inc., Philadelphia,
PA, USA).
5. Peristaltic pump (Sartorius Stedim Biotech GmbH, Go ¨ttingen,
Germany) with Tandem 1082 pump head, with a Masterflex
Pharmed #15 (Cole Parmer GmbH, Wertheim, Germany)
tube and transparent silicon tubes (VWR International, Radnor, PA, USA) with sterile Luer lock connections (Sarstedt,
Nu ¨mbrecht, Germany).
6. Opticap XL 1 Capsule with Polygard
® -CR 5.0 μm (Nominal)—sterile (Merck KGaA, Darmstadt, Germany).
7. 1 L PBS—Sterile (#L1825, Biochrom, Berlin, Germany).
8. Sodium hydroxide (Carl Roth GmbH, Karlsruhe, Germany).
9. Sterile bottles (0.5 L, Schott AG, Mainz, Germany).
10. Sterile workbench (ESCO Infinity Class II, Esco Lifesciences
GmbH, Friedberg, Germany).
2.2.2 Preparation
of Buffers and Membranes
1. PBS (#L1825, Biochrom, Berlin, Germany).
2. Deionized water (Milli-Q Academic, Q-Gard
® 1, Merck
KGaA, Darmstadt, Germany).
3. PEG 8000 (biotechnology grade, VWR International, Radnor,
PA, USA).
4. Sodium chloride (Sigma-Aldrich, Hamburg, Germany).
5. Sodium hydroxide 1 M NaOH (Carl Roth GmbH, Karlsruhe,
Germany).
6. 100-mL, 250-mL, and 1-L glass beakers (Carl Roth GmbH,
Karlsruhe, Germany).
7. 100-mL, 250-mL, and 500-mL measuring cylinders (Carl
Roth GmbH, Karlsruhe, Germany).
8. Analytical scale (Kern & Sohn GmbH, Balingen-Frommern,
Germany).
9. Magnetic stirring bar (Carl Roth GmbH, Karlsruhe,
Germany).
Viral Nanoplex Vaccine Production
227