The Subtle Relationship Between Cells and Bubbles, How Does BioLink

With the booming development of biopharmaceutical industry, antibody drugs, gene therapy and cell therapy are getting more and more attention. Bioreactor is one of the core equipment in biotechnology, as the “incubator” of cells, its stable and precise control of key parameters affecting cell growth and product quality is of particular concern.

Aeration, as one of the ways for the “incubator” to provide key nutrients to the cells, is a parameter of great concern in large-scale cell culture, and has a huge impact on the microenvironment of cell culture: it affects dissolved oxygen mass transfer, pCO2, bubbles, and so on, and improper ventilation methods can cause cell damage. The larger the bubbles, the worse the oxygen transfer capacity; conversely, the better the oxygen transfer capacity. Meanwhile, Tan Wensong et al. had studied the aeration rate and bubble diameter on cell damage, and found that the higher the aeration rate and the smaller the bubble diameter, the stronger the killing effect, and the killing effect of stirring shear force is seriously dependent on aeration. Therefore according to the diameter of the bubbles entering the cell culture bags, aeration and agitation must be strictly controlled in the appropriate range.

Typically, bubbles travelling from the gas distributor to the outlet create three areas of high shear force: the outlet of the gas distributor, the bubble rise area and the bubble burst area. How to get cells and bubbles to “coexist harmoniously” in these areas is particularly important.

In order to balance agitation, aeration and cell growth, by optimising the mixing paddles and gas distributor structure, BioLink has developed and launched the CytoLinX^® BR series of single-use bioreactor systems and CytoLinX^® BR single-use down-stirred bioreactor bags including microbubble, 0.3mm medium bubble and 0.8mm large bubble gas distributor. The combination of gas distributor can be freely matched according to the user's needs to achieve excellent performance.

Based on the CytoLinX^® BR series single-use bioreactor system and CytoLinX^® BR single-use down-stirred bioreaction bag, CHO cells were cultured in 50L, 200L and 2000L volumes respectively using different combinations of gas distributors and conventional perfusion culture in order to achieve excellent cell proliferation, cell viability and good metabolite control. The results are shown in Figure 1- Figure 3.

Figure 1

CHO K1 cells were cultured using conventional flow-addition culture based on the CytoLinX^® BR 50L single-use bioreactor system and the CytoLinX^® BR 50L single-use down-stirred bioreactor bag. The reaction bag was configured with a BioLink 0.3mm medium bubble gas distributor. Cells proliferated rapidly and maintained good cell activity, with lactate and ammonium concentrations consistently maintained at low levels (lactate and ammonium concentrations are not listed).

Figure 2

CHO DG44 cells were cultured using conventional perfusion culture based on the CytoLinX^® BR 50L single-use bioreactor system and CytoLinX^® BR 50L single-use down-stirred bioreactor bag. The reaction bag was configured with a BioLink 0.3mm medium bubble gas distributor. Cells proliferated rapidly and maintained good cell activity with lactate and ammonium concentrations consistently maintained at low levels (lactate and ammonium concentrations are not listed).

Figure 3

CHO K1 cells were cultured using conventional perfusion culture based on the CytoLinX^® BR 2000L single-use bioreactor system and the CytoLinX^® BR 2000L single-use down-stirred bioreactor bag. The reaction bag was equipped with a BioLink 0.8 mm large bubble gas distributor. Cells proliferated rapidly and maintained good cellular activity with consistently low lactate and ammonium concentrations (lactate and ammonium concentrations are not listed).

Conclusion

The results show that the optimised CytoLinX^® BR series of single-use bioreactor systems with CytoLinX^® BR single-use down-stirred bioreactor bags configured with different combinations of gas distributors exhibit excellent process control in different volumes, and the cells maintain excellent growth and viability. It provides a reliable and efficient single-use solution for the scale-up and large-scale production of recombinant proteins and antibodies.

BioLink CytoLinX^® BR Series Single-Use Tank Bioreactor

                                                                                              CytoLinX^® BR Single-use Stirred Tank Bioreactor

CytoLinX^® BR offers the following advantages:

►A single control cabinet can control several reactors which can realise a significant cost reduction;

►Stable PCS 7 system, in line with ISA 88 control standard, which can achieve the whole plant control;

►Software design, in accordance with 21 CFR Part 11, has a user-friendly interface;

►Flexible configuration, can be partially customised according to user requirements;

►Bioprocess equipment accessories are imported high-end brands, and have passed factory testing to ensure normal use;

►Consumable spargers are available in large bubble, medium bubble, and microbubble sizes to meet a variety of process needs

►Provide perfect FAT, SAT, IQ, OQ documents and services;

►With rich application and operation support, and high speed response;

With the BioLinX^® WB series of rocking bioreactors of BioLink, it is possible to achieve infinite scale-up production, which is an excellent choice for optimising process single-use equipment.

BioLink CytoLinX^® WB Series Single-Use Rocking Bioreactors

                                                                                          CytoLinX^® WB Single-use Wave Bioreactor

Main characteristics:

● Easy to operate software interface with stable operation and audit trail function;

● 10L, 20L and 50L tray specifications are available. The installation of tray and culture bag does not need to remove the screws, which can be installed simply and quickly;

● Support perfusion function, the flow rate of the pump has an automatic calibration function to achieve accurate replenishment and harvesting;

● Ventilation, pH/DO, perfusion function modules can be easily combined and selected according to process requirements;

● Support DO associated N2 reverse regulation, be applicable to a wide range of fields to help cell therapy customers more accurately control cell state;

● The system can be regulated by frequency conversion and swing mode, which can further reduce shear force and make cell culture more gentle;

● Use gas mass flow controller (MFC) to measure the gas accurately;

●The whole machine is equipped with monitoring alarm and protection functions, which can automatically sense and diagnose faults and alarms;

● Multiple bags are available, including basic type, pH & DO electrodes type, and perfusion type;

● Rich application and operation support, and high response speed;

● Fast delivery and flexible customization possibilities.

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