6. Sterile single-use pipette tips (e.g., serological pipettes, Sarstedt, Nu ¨mbrecht, Germany).
7. Pipette holder/rubber bulb (e.g., pipetus, Hirschmann Laborger€ ate, Eberstadt, Germany).
8. 6-well tissue-culture plates (e.g., Sarstedt, Nu ¨mbrecht,
Germany).
9. Distilled water, cell-culture grade, sterile (e.g., Milli-Q Academic, Millipore, Burlington, Massachusetts, USA).
10. 100 mL glass bottle, sterile (e.g., Glasger€ atebau Ochs, Bovenden, Germany).
11. Water bath (e.g., Memmert, Schwabach, Germany).
12. Microwave.
13. Incubator (e.g., KB 115 E 3.1, Binder, Tuttlingen, Germany).
2.6.3 Materials
for the Determination
of the 50% Tissue Culture
Infective Dose (TCID 50 )
1. Spodoptera frugiperda cell suspension with a cell density of
1.0 Â 10
5 cells/mL (e.g., Sf-9 Insect Cells, Merck, Darmstadt,
Germany).
2. Baculovirus stock P2 (e.g., AcMNPV-Orf2-PCV2).
3. Insect cell culture medium (e.g., Sf900 II, Thermo Fisher
Scientific, Waltham, MA, USA).
4. 96-well plates (e.g., TC Plate 96, Sarstedt, Nu ¨mbrecht,
Germany).
5. Reagent reservoir (e.g., VWR, Radnor, Pennsylvania, USA).
6. Multichannel pipette (e.g., Research Plus, Eppendorf, Hamburg, Germany).
7. Single-use pipette reservoirs (e.g., 25-mL pipette reservoir,
Argos, Vernon Hills, IL, USA).
8. Single-use pipette tips (e.g., Sarstedt, Nu ¨mbrecht, Germany).
9. Incubator (e.g., KB 115 E 3.1, Binder, Tuttlingen, Germany).
2.7 Materials
for Infection of Sf-cells
with Baculovirus
and Protein Production
in a Stirred-Tank
Bioreactor
In this part of the chapter, the materials for the infection of insect
cells, using BEVS for protein production in a stirred-tank bioreactor, and the harvest of those produced recombinant proteins, are
described.
2.7.1 Materials
for the Infection of Sf-cells
with Baculovirus
1. Spodoptera frugiperda cell suspension (1 L) with a cell density
of 1 Â 10
6 cells/mL (e.g., TriEx Sf-9, Merck, Darmstadt,
Germany).
2. Insect cell culture medium (e.g., Sf900 II, Thermo Fisher
Scientific, Waltham, MA, USA).
Antigen Production in Insect Cells
105
7. Pipette holder/rubber bulb (e.g., pipetus, Hirschmann Laborger€ ate, Eberstadt, Germany).
8. 6-well tissue-culture plates (e.g., Sarstedt, Nu ¨mbrecht,
Germany).
9. Distilled water, cell-culture grade, sterile (e.g., Milli-Q Academic, Millipore, Burlington, Massachusetts, USA).
10. 100 mL glass bottle, sterile (e.g., Glasger€ atebau Ochs, Bovenden, Germany).
11. Water bath (e.g., Memmert, Schwabach, Germany).
12. Microwave.
13. Incubator (e.g., KB 115 E 3.1, Binder, Tuttlingen, Germany).
2.6.3 Materials
for the Determination
of the 50% Tissue Culture
Infective Dose (TCID 50 )
1. Spodoptera frugiperda cell suspension with a cell density of
1.0 Â 10
5 cells/mL (e.g., Sf-9 Insect Cells, Merck, Darmstadt,
Germany).
2. Baculovirus stock P2 (e.g., AcMNPV-Orf2-PCV2).
3. Insect cell culture medium (e.g., Sf900 II, Thermo Fisher
Scientific, Waltham, MA, USA).
4. 96-well plates (e.g., TC Plate 96, Sarstedt, Nu ¨mbrecht,
Germany).
5. Reagent reservoir (e.g., VWR, Radnor, Pennsylvania, USA).
6. Multichannel pipette (e.g., Research Plus, Eppendorf, Hamburg, Germany).
7. Single-use pipette reservoirs (e.g., 25-mL pipette reservoir,
Argos, Vernon Hills, IL, USA).
8. Single-use pipette tips (e.g., Sarstedt, Nu ¨mbrecht, Germany).
9. Incubator (e.g., KB 115 E 3.1, Binder, Tuttlingen, Germany).
2.7 Materials
for Infection of Sf-cells
with Baculovirus
and Protein Production
in a Stirred-Tank
Bioreactor
In this part of the chapter, the materials for the infection of insect
cells, using BEVS for protein production in a stirred-tank bioreactor, and the harvest of those produced recombinant proteins, are
described.
2.7.1 Materials
for the Infection of Sf-cells
with Baculovirus
1. Spodoptera frugiperda cell suspension (1 L) with a cell density
of 1 Â 10
6 cells/mL (e.g., TriEx Sf-9, Merck, Darmstadt,
Germany).
2. Insect cell culture medium (e.g., Sf900 II, Thermo Fisher
Scientific, Waltham, MA, USA).
Antigen Production in Insect Cells
105