1. The BIOne benchtop single-use bioreactor system for mammalian
cell growth and recombinant protein production as a
robust model for bioprocess development
Distek, Inc. has developed a benchtop scale single-use bioreactor (SUB) system for mammalian cell growth and recombinant protein production. The pre-sterilized BIOne
system is engineered with a disposable headplate welded to a triple-layered liner that can be easily inserted into a non-sterile bioreactor glass vessel, converting it to a sterile,
disposable SUB within a matter of seconds. The Distek BIOne system significantly reduces turnaround time by allowing users to seamlessly transition to a disposable platform
while utilizing their existing capital equipment, without compromising the scalability of their current process. In this study, mammalian cultures were conducted by using
either the Distek BIOne system or the traditional, non-disposable bioreactor system. Comparable cell culture performance was observed supporting that the Distek BIOne
system provides a robust model for bioprocess development.
Benchtop scale bioreactors provide a cost efficient model for process development and characterization that can be readily scaled-up or scaled-down from production scale.
Given that upstream process development and characterization are rigorous and complicated, high-throughput is vital to ensure efficiency and avoid delays. Thus, many have
utilized the single-use platform to significantly reduce turnaround time by eliminating cleaning, assembling, and autoclaving. Distek Inc. has developed the BIOne benchtop
single-use bioreactor (SUB) system with an innovative liner technology that inserts into an existing glass vessel to easily convert the existing non-disposable platform to a
disposable, single-use platform. Upon media addition, the triple-layered liner expands and molds to the glass vessel, preserving the geometry of the existing bioreactor. The
BIOne is fully compatible with your existing equipment including controller, probes and agitator, thus, no new capital investment required.
This study evaluated the suitability of the BIOne system for mammalian cell growth and recombinant protein production. Relative to a glass vessel bioreactor (control), three
aspects of comparability were assessed: (1) Volumetric Mass Transfer Coefficient (kL
a) of Oxygen, (2) Process Control and (3) Growth Kinetics and Titer. Volumetric mass
transfer coefficient (kL
a) of oxygen is a critical parameter in evaluating the efficiency with which a bioreactor can meet the oxygen demands of a bioprocess. kL
a was assessed
over a range of agitation rates to verify the BIOne’s ability to achieve similar values relative to a glass vessel bioreactor. Process control capabilities were assessed to determine
whether bioreactor setpoints could be effectively maintained over the course of a run. Growth kinetics and final titer were evaluated to establish the BIOne’s suitability for
mammalian cell culture applications. Relative to a glass vessel bioreactor, similar performance attributes in these key areas would demonstrate the BIOne’s utility as a robust
model for bioprocess development.
Mark C. Arjona and Michael Lau Ph.D., Distek, Inc. - North Brunswick, New Jersey
ABSTRACT
INTRODUCTION
We would like to thank Dr. Sarwat Khattak and her team at BTEC of NC State University for their cell culture data.
Presented at BioProcess International Conference & Exposition, October 5-7, 2016 (Boston, Massachusetts, USA)
METHOD AND RESULTS
Data was generated in a BIOne SUB and a non-disposable, glass vessel bioreactor using identical process parameters and equipment. Process control and cell culture were
evaluated in a chemically-defined, fed-batch CHO process. Titer was quantified once culture viability reached ≥80%.
CONCLUSION
1. The BIOne system is a suitable benchtop-scale SUB
for mammalian cell growth and recombinant
protein production, effectively eliminating the time
and costs associated with cleaning and autoclaving
non-disposable bioreactor vessels.
2. The BIOne is capable of achieving similar kL
a values
as non-disposable, glass vessel bioreactors when
run with identical parameters.
3. Utilizing the BIOne did not affect process control
capabilities (%DO, pH, temperature and agitation)
over the course of the culture run.
4. Similar cell growth profiles and protein production
were achieved in the BIOne system relative to the
non-disposable, glass vessel bioreactor.
Figures 6 thru 8 - CHO growth and protein production. Similar growth profiles and protein production were observed between the BIOne and control. Performance results
indicate that the BIOne system is a suitable benchtop-scale SUB for CHO growth and protein production.
GROWTH KINETICS & TITER
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
10.0
0 1 2 3 4 5 6 7
ViableCellDensity
(x106cells/mL)
Day
BIOne
Control
Figure 6. Viable Cell Density
0
10
20
30
40
50
60
70
80
90
100
0 1 2 3 4 5 6 7
Viability
(%)
Day
BIOne
Control
Figure 7. Viability
0.0
0.2
0.4
0.6
0.8
1.0
1.2
BIOne Control
NormalizedTiter
Figure 8. Normalized Titer
Figures 2 thru 5 - Process control. Similar % dissolved oxygen (%DO) and pH were observed between the BIOne and control. BIOne effectively maintained temperature and
agitation setpoints. Utilizing the BIOne does not adversely affect process control capabilities.
Figure 1. Volumetric mass transfer coefficient (kL
a) of oxygen. kL
a was evaluated via static gassing-out method. Similar kL
a values were observed in the BIOne and glass
vessel control.
PROCESS CONTROL
VOLUMETRIC MASS TRANSFER COEFFICIENT (kL
a) of OXYGEN
0
20
40
60
80
100
120
0 1 2 3 4 5 6 7
DissolvedOxygen
(%)
Day
BIOne DO Control DO
Setpoint = 50%
Figure 2. Dissolved Oxygen
6.0
6.2
6.4
6.6
6.8
7.0
7.2
7.4
7.6
7.8
8.0
0 1 2 3 4 5 6 7
pH
Day
BIOne pH Control pH
Setpoint = pH 7.0
Deadband = 6.70 - 7.30
Setptt oint = pH 7.0
Figure 3. pH
31
33
35
37
39
41
43
0 24 48 72 96 120 144 168
Temperature
(°C)
Time (hours)
BIOne Temperature
Setpoint = 37°C
Figure 4. BIOne Temperature
0
25
50
75
100
125
150
175
200
225
250
0 24 48 72 96 120 144 168
AgitaƟon
(rpm)
Time (hours)
BIOne AgitaƟon
Setpoint = 125 rpm
Figure 5. BIOne Agitation
GLASS
VESSEL
SINGLE-USE
LINER
SINGLE-USE
HEADPLATE
2.0
1.7
2.0
2.4
4.1
3.8
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
3L Glass 3L Glass 3L Glass
Vessel Vessel Vessel
150 rpm 300 rpm 450 rpm
2L BIOne 2L BIOne 2L BIOne
kLa(hr-1)
Figure 1. kL
a
Culture Vessels 2L BIOne 3L Glass Vessel
Working Volume 1.7L
Temperature 37°C
Overlay None
Sparger Flow Rates Air @ 0.05 vvm
Agitation 150, 300 & 450 RPM
Medium Water
Sparger 7-Hole, Flute Sparger
Impeller 45 mm Pitch Blade (Vortex Orientation)
BIOne
SINGLE-USE HEADPLATE