The document summarizes a study that evaluated the Distek BIOne benchtop single-use bioreactor system for mammalian cell culture and recombinant protein production. The key findings were:
1. The BIOne system achieved similar volumetric mass transfer coefficients (kLa) of oxygen as a traditional glass vessel bioreactor when run at identical parameters.
2. The BIOne effectively maintained process control parameters like dissolved oxygen, pH, temperature, and agitation at their setpoints over the culture period, comparable to the glass vessel bioreactor.
3. Mammalian cell growth profiles and final protein titers were similar when cultures were run in the BIOne versus the glass vessel bi
Scalability of Cell Culture Processes in Single-use Bioreactors using Differe...KBI Biopharma
Niket Bubna, Cameron T. Phillips, Sigma S. Mostafa and AbhinavA. Shukla. KBI Biopharma, Durham, NC
253rd ACS National Meeting & Exposition
April 2-6, 2017 • San Francisco, CA
#acsSanFran • www.acs.org/SanFran2017
Next Generation Recombinant Protein ManufacturingKBI Biopharma
Next Generation Processes: What Model Works Best to Manufacture Recombinant Proteins in Asia?
BioPharma Asia 2017
Suntec Convention Center. Singapore, March 22, 2017
Thomas Jung, M.S. Vice President, Business Development
KBI Biopharma Inc.
Scalability of Cell Culture Processes in Single-use Bioreactors using Differe...KBI Biopharma
Niket Bubna, Cameron T. Phillips, Sigma S. Mostafa and AbhinavA. Shukla. KBI Biopharma, Durham, NC
253rd ACS National Meeting & Exposition
April 2-6, 2017 • San Francisco, CA
#acsSanFran • www.acs.org/SanFran2017
Next Generation Recombinant Protein ManufacturingKBI Biopharma
Next Generation Processes: What Model Works Best to Manufacture Recombinant Proteins in Asia?
BioPharma Asia 2017
Suntec Convention Center. Singapore, March 22, 2017
Thomas Jung, M.S. Vice President, Business Development
KBI Biopharma Inc.
Growth Characteristics Modeling of Lactobacillus acidophilus using RSM and ANNGanga Sahay Meena
The culture conditions viz. additional carbon and n
itrogen content, inoculum size, age, temperature an
d pH of
Lactobacillus acidophilus
were optimized using response surface methodology (
RSM) and artificial neural network
(ANN). Kinetic growth models were fitted to cultiva
tions from a Box-Behnken Design (BBD) design experi
ments for
different variables. This concept of combining the
optimization and modeling presented different optim
al conditions
for
L. acidophilus
growth from their original optimization study. Thr
ough these statistical tools, the product yield
(cell mass) of
L. acidophilus
was increased. Regression coefficients (R
2
) of both the statistical tools predicted that
ANN was better than RSM and the regression equation
was solved with the help of genetic algorithms (GA
). The
normalized percentage mean squared error obtained f
rom the ANN and RSM models were 0.06 and 0.2%,
respectively. The results demonstrated a higher pre
diction accuracy of ANN compared to RSM
Single-Use-Bioreactors-A-Comprehensive-Examination.
Single-use bioreactors (SUBs) have revolutionized biopharmaceutical production, offering advantages over traditional bioreactors.
Growth Characteristics Modeling of Lactobacillus acidophilus using RSM and ANNGanga Sahay Meena
The culture conditions viz. additional carbon and n
itrogen content, inoculum size, age, temperature an
d pH of
Lactobacillus acidophilus
were optimized using response surface methodology (
RSM) and artificial neural network
(ANN). Kinetic growth models were fitted to cultiva
tions from a Box-Behnken Design (BBD) design experi
ments for
different variables. This concept of combining the
optimization and modeling presented different optim
al conditions
for
L. acidophilus
growth from their original optimization study. Thr
ough these statistical tools, the product yield
(cell mass) of
L. acidophilus
was increased. Regression coefficients (R
2
) of both the statistical tools predicted that
ANN was better than RSM and the regression equation
was solved with the help of genetic algorithms (GA
). The
normalized percentage mean squared error obtained f
rom the ANN and RSM models were 0.06 and 0.2%,
respectively. The results demonstrated a higher pre
diction accuracy of ANN compared to RSM
Single-Use-Bioreactors-A-Comprehensive-Examination.
Single-use bioreactors (SUBs) have revolutionized biopharmaceutical production, offering advantages over traditional bioreactors.
High-throughput Miniaturized Bioreactors for Cell Culture Process Developmen...KBI Biopharma
Decreasing the timeframe for cell culture process development has been a key goal towards accelerating biopharmaceutical development. Automated Micro-scale Bioreactors (ambrTM) is an advanced micro bioreactor system with miniature single-use bioreactors with a 9-15mL working volume controlled by an automated workstation. This system was compared to conventional bioreactor systems in terms of its performance for the production of a monoclonal antibody and a non-antibody molecule in recombinant Chinese Hamster Ovary (CHO) cell lines.
The miniaturized bioreactor system was found to produce cell culture profiles that matched across scales to 3L, 15L and 200L stirred tank bioreactors. Moreover, changes to important process parameters in ambrTM resulted in predictable cell growth, viability and titer changes, which were in good agreement to historical data from the larger scales. ambrTM was found to successfully reproduce variations in temperature, dissolved oxygen and pH conditions similar to the larger bioreactor systems. Additionally, the miniature bioreactors were found to react well to perturbations in pH and dissolved oxygen through adjustments to the PID control loop. Overall, the studies demonstrate the utility of the ambrTM system as a high throughput system for cell culture process development.
Scalability of a Single-use Bioreactor Platform for Biopharmaceutical Manufac...KBI Biopharma
Increasing adoption of single-use technologies for bioprocessing along with higher titers from cell culture bioreactor processes has allowed clinical and even commercial manufacturing to be successfully performed in 2000 L-scale single-use bioreactors. Several biopharmaceutical manufacturers have successfully adopted single-use bioreactors for production. However, information about process scalability from glass bioreactors to 2000 L single-use bioreactors for different types of CHO cell lines is not widely available. Here we provide an overview of the key
differences between single-use and conventional stainless steel bioreactors, and highlight factors that are employed while scaling-up from small-scale glass bioreactors to 2000 L-scale single-use bioreactors. Several case studies focusing on process performance across scales into single-use bioreactors are provided. This analysis confirms that the 2000 L-scale single-use bioreactorsystem can be robustly employed for biopharmaceutical manufacturing.
A bioreactor is a type of fermentation vessel that is used for the production of various chemicals and biological reactions. It is a closed container with adequate arrangement for aeration, agitation, temperature and pH control, and drain or overflow vent to remove the waste biomass of cultured microorganisms along with their products.
Integration of Cell Line and Process Development to Expedite Delivery of Bisp...KBI Biopharma
Authored and Presented by: Dane A. Grismer, Yogender K. Gowtham, Srivatsan Gopalakrishnan, David. W. Chang,
Niket Bubna, Ph.D., and Sigma S. Mostafa, Ph.D.
Industry trends are moving toward the implementation of automated PAT strategies designed to bring analytics closer to the operation, eliminate gaps in real-time process monitoring and develop a more rapid, deeper understanding of the bioprocess.
Integrated utilization of high-throughput bioreactors & high-throughput analy...KBI Biopharma
There is a strong impetus towards rapidly advancing an increasing number of novel biotherapeutics to clinical trials. However, development of cell culture processes is labor intensive and time consuming. KBI focuses on a high throughput process development (HTPD) approach using high-throughput miniaturized bioreactors and high throughput analytics that generate growth, productivity and product quality data that match those seen with classical systems. This approach enables a significant reduction in the cell culture process development timeline and costs for investigational biopharmaceuticals to reach the clinic.
High Throughput Bioreactor Mimetic in Early and Late Stage Process DevelopmentKBI Biopharma
A presentation by KBI Scientist Shahid Rameez, Ph.D. at the American Chemical Society Annual Meeting– Biochemical Technology (BIOT) Division, New Orleans, LA
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