l
Sterile glass bottle topped with multiport connection system for
bioreactors (Schott, Mainz, GER): one air filter (Merck Millipore, Burlington, MA, USA), one connector to probe vent of
bioreactor (e.g., Luer lock [Sigma-Aldrich, St. Louis, MO,
USA]), additional connectors should be sealed.
l
PBS (Biochrom, Berlin, GER), adjusted to pH 6.4 with 1 M
HCl (Carl Roth, Karlsruhe, GER), and filtered through 0.2 μm
bottle-top filter (Fisher Scientific, Hampton, NH, USA). All
buffers should be sterilized in an autoclave, or using a bottletop filter.
l
Water, 0.2 μm filtered, sterilized.
l
1 M NaOH (Carl Roth, Karlsruhe, GER), 0.2 μm filtered.
l
Serological pipettes and holder (Sarstedt, Nu ¨mbrecht, GER).
l
Sterile pipette tips and pipette (Eppendorf, Hamburg, GER).
l
1.5 mL tubes (Eppendorf, Hamburg, GER).
l
15 mL and/or 50 mL centrifuge tube (Sarstedt,
Nu ¨mbrecht, GER).
2.3.4 BV Purification via
TFF
l
Sartocon
® Slice 200 Polyethersulfone (PES) membrane (cassette), MWCO (molecular weight cut off) 300 kDa, filtration
area 200 cm
2 (Sartorius, Go ¨ttingen, GER) (see Note 5).
l
Sartocon
® Slice 200 Holder (Sartorius, Go ¨ttingen, GER).
l
PBS (Merck Millipore, Burlington, MA, USA), adjusted to
pH 6.4 with 1 M HCl (Carl Roth, Karlsruhe, GER). 0.2 μm
filtered, sterilized.
l
Water, 0.2 μm filtered, sterilized.
l
1 M NaOH (Carl Roth, Karlsruhe, GER), 0.2 μm filtered.
l
Serological pipettes (Sarstedt, Nu ¨mbrecht, GER) and holder.
l
Sterile pipette tips and pipette (Eppendorf, Hamburg, GER).
l
1.5 mL tubes (Eppendorf, Hamburg, GER).
l
15 mL and/or 50 mL centrifuge tube (Sarstedt,
Nu ¨mbrecht, GER).
Regeneration of Sartocon
® Slice 200
l
1 M NaOH (Carl Roth, Karlsruhe, GER), 0.2 μm filtered.
l
Water, 0.2 μm filtered, sterilized.
l
Dry-heat oven (Memmert, Schwabach, GER), 50
C.
l
20% ethanol (Carl Roth, Karlsruhe, GER), 0.2 μm filtered.
374
Friederike Eilts et al.
Sterile glass bottle topped with multiport connection system for
bioreactors (Schott, Mainz, GER): one air filter (Merck Millipore, Burlington, MA, USA), one connector to probe vent of
bioreactor (e.g., Luer lock [Sigma-Aldrich, St. Louis, MO,
USA]), additional connectors should be sealed.
l
PBS (Biochrom, Berlin, GER), adjusted to pH 6.4 with 1 M
HCl (Carl Roth, Karlsruhe, GER), and filtered through 0.2 μm
bottle-top filter (Fisher Scientific, Hampton, NH, USA). All
buffers should be sterilized in an autoclave, or using a bottletop filter.
l
Water, 0.2 μm filtered, sterilized.
l
1 M NaOH (Carl Roth, Karlsruhe, GER), 0.2 μm filtered.
l
Serological pipettes and holder (Sarstedt, Nu ¨mbrecht, GER).
l
Sterile pipette tips and pipette (Eppendorf, Hamburg, GER).
l
1.5 mL tubes (Eppendorf, Hamburg, GER).
l
15 mL and/or 50 mL centrifuge tube (Sarstedt,
Nu ¨mbrecht, GER).
2.3.4 BV Purification via
TFF
l
Sartocon
® Slice 200 Polyethersulfone (PES) membrane (cassette), MWCO (molecular weight cut off) 300 kDa, filtration
area 200 cm
2 (Sartorius, Go ¨ttingen, GER) (see Note 5).
l
Sartocon
® Slice 200 Holder (Sartorius, Go ¨ttingen, GER).
l
PBS (Merck Millipore, Burlington, MA, USA), adjusted to
pH 6.4 with 1 M HCl (Carl Roth, Karlsruhe, GER). 0.2 μm
filtered, sterilized.
l
Water, 0.2 μm filtered, sterilized.
l
1 M NaOH (Carl Roth, Karlsruhe, GER), 0.2 μm filtered.
l
Serological pipettes (Sarstedt, Nu ¨mbrecht, GER) and holder.
l
Sterile pipette tips and pipette (Eppendorf, Hamburg, GER).
l
1.5 mL tubes (Eppendorf, Hamburg, GER).
l
15 mL and/or 50 mL centrifuge tube (Sarstedt,
Nu ¨mbrecht, GER).
Regeneration of Sartocon
® Slice 200
l
1 M NaOH (Carl Roth, Karlsruhe, GER), 0.2 μm filtered.
l
Water, 0.2 μm filtered, sterilized.
l
Dry-heat oven (Memmert, Schwabach, GER), 50
C.
l
20% ethanol (Carl Roth, Karlsruhe, GER), 0.2 μm filtered.
374
Friederike Eilts et al.