Facing the Challenges in Vaccine Upstream Bioprocessing - 31

Facing the Challenges in Vaccine Upstream Bioprocessing * Vero Perfusion, Packed-Bed Vessels Intensify Vaccine Production

In this study, we cultivated Vero cells in perfusion mode in Eppendorf BioBLU® 5p Single-Use
Vessels with a built-in basket impeller prepacked
with Fibra-Cel (Figure 1). We achieved a high
cell density of approximately 43 million cells/
mL, demonstrating great potential for Vero
cell-based vaccine production using Fibra-Cel
packed-bed vessels.

Materials and Methods
Cell Line and Medium
We cultivated adherent Vero cells (ATCC, CCL81TM) in Dulbecco's modified Eagle's medium
(DMEM) (Thermo Fisher Scientific), supplemented with 1× Antibiotic-Antimycotic (Thermo
Fisher Scientific) and 1% (v/v) heat-inactivated
fetal bovine serum.

Figure 1. BioBLU 5p Single-Use Vessel

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which is equivalent to about 212 roller bottles, or
29 10-layer, stacked-plate, cell culture vessels.2

Bioprocess Control
The bioprocess was controlled with a BioFlo®
320 bioprocess control station. The BioBLU 5p
Single-Use Vessel was inoculated with 530 mL of
inoculum, which had a cell density of 3.0 × 106
cells/mL and a cell viability of 99%, resulting in an
inoculation density of 4.6 × 105 cells/mL. The cells
were cultivated at 37°C.

Growth Surface

Dissolved oxygen (DO) was measured using a
polarographic ISM® sensor (Mettler Toledo). DO
was controlled at 50% by automatic gassing at a
flow of 0.002 - 0.5 SLPM in the 3-Gas Auto mode
via a macrosparger. To reduce foaming caused by
high gas flow in the late culture stage, we limited
air flow to 0.002-0.2 SLPM and oxygen flow to
0-0.5 SLPM.

We cultivated Vero cells in a BioBLU 5p SingleUse Vessel with a working volume of 3.5 L. Each
BioBLU 5p contains 150 g of Fibra-Cel disks.
These provide a growth surface of 180,000 cm2,

The BioBLU 5p Single-Use Vessel was equipped
with an optical pH sensor and the pH was
controlled at 7.1 (deadband = 0.1) via a cascade

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Facing the Challenges in Vaccine Upstream Bioprocessing

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