The BIOne Benchtop Single-Use Bioreactor System For Mammalian Cell Growth And Recombinant Protein Production As A Robust Model For Bioprocess Development

• Distek, Inc.

Abstract

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.

Introduction

Benchtop scale bioreactors provide a cost efficient model for process development and characterization that can be readily scaledup 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 (kLa) of Oxygen, (2) Process Control and (3) Growth Kinetics and Titer. Volumetric mass transfer coefficient (kLa) of oxygen is a critical parameter in evaluating the efficiency with which a bioreactor can meet the oxygen demands of a bioprocess. kLa 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.

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%.

Volumetric Mass Transfer Coefficient (k_La) of Oxygen

Table 1.

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

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 2. Dissolved Oxygen

Figure 3. pH

Figure 4. BIOne Temperature

Figure 5. BIOne Agitation

Growth Kinetics & Titer

Figures 6 thru 8 - CHO growth and protein production. Similar growth profiles and protein production were observed between the BIOne and control. Performance results non-disposable, glass vessel bioreactor. indicate that the BIOne system is a suitable benchtop-scale SUB for CHO growth and protein production.

Figure 6. Viable Cell Density

Figure 7. Viability

Figure 8. Normalized Titer

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 kLa values as nondisposable, 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 nondisposable, glass vessel bioreactor.

Acknowledgement

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)