Optimization of Antibody Scale-Up Production Based on Single-Use Technology

In order to further reduce costs and increase efficiency, an increasing number of pharmaceutical companies are adopting the process of pre-cultivating N-1 generation seed in N vessels. This article introduces the process setting of small-scale cultivation using BioLink's single-use bioprocess technology and compares its advantages with traditional batch cultivation. It provides a more flexible process solution selection.

The CytoLinX® WB single-use wave bioreactor and CytoLinX® BR 50-2000L series single-use bioreactor from BioLink can both achieve seamless scale-up production, providing a powerful single-use equipment solution for process optimization.

Equipment Advantages


CytoLinX® WB single-use wave bioreactor

The characteristics of CytoLinX® WB single-use wave bioreactor include:

  • Cultivation volume ranges from 0.1 L to 100 L

  • User-friendly and stable software interface, compliant with 21 CFR Part 11 requirements

  • Optional pH and DO modules

  • Precise gas measurement using gas mass flow controllers (MFC)

  • Diverse range of consumables, including basic, pH & DO electrode, and perfusion options

  • Covers consumable designs for cell therapy applications


CytoLinX® BR single-use tank bioreactor

The characteristics of CytoLinX® WB single-use bioreactor include:

  • The cultivation volume of the reactor ranges from 50 L to 2000 L

  • User-friendly interface with "plug and play" functionality and "1 control multiple" capability

  • Utilizes a stable DCS control system that complies with the ISA 88 control standard

  • Software design compliant with 21 CFR Part 11

  • Various consumable options available, including bottom sparger with large bubbles, medium bubbles, and microbubbles, to meet different process requirements

The effectiveness of the CytoLinX® BR single-use bioreactor and CytoLinX® WB single-use wave bioreactor was evaluated in optimizing antibody production using a Fed-Batch process. In this experiment, CHO-K1 and CHO-DG44 cells were used. Cell expansion was carried out in the CytoLinX® WB single-use wave bioreactor, followed by transferring the cells to a 200L CytoLinX® BR single-use bioreactor for cultivation. Both experiments resulted in the expected antibody yields.

The following image demonstrates the excellent control effect of process parameters exhibited by the CytoLinX® BR single-use bioreactor during cell culture.


▲ Figure 1: CytoLinx® BR-200 Results of a 14-day cell culture (curves represent weight, DO, temperature, and pH)

Experimental Method:

In this experiment, CHO-DG44 and CHO-K1 cells were used. The cells were revived from frozen stocks according to standard methods and then passaged and amplified in shake flasks every three to four days. Once the cell count was sufficient, the cells were transferred to CytoLinX® WB and CytoLinX® BR-200 bioreactors for further cultivation.


Experiment 1: CHO-DG44 Cell Expansion, N-1 Expansion, and Perfusion Culture Stage Operation Parameters and Conditions

Result of Experiment 1:  The cell growth curve and biochemical data are shown in Figure 2 and Figure 3, respectively.

experimental-method-7.jpg▲ Figure 2: CHO DG44 Growth density and viability curve graph of cells

Figure 3: CHO-DG44 Biochemical data of cells: lactate and pCO2 curve graph

experimental-method-8.jpgExperiment 2: Operational parameters and conditions for CHO-K1 cell expansion, N-1 expansion stage, and fed-batch cultivation stage

Result of Experiment 2: Cell growth curve and biochemical data of cell culture are shown in Figure 4 and Figure 5.

Figure 4: CHO- K1 Growth density and viability curve graph of cells

Figure5: CHO-K1 Biochemical data of cells: lactate and pCO2 curve graph


The initial culture volume of CytoLinX®-200 is 140 L. With daily supplementation, the final volume of CytoLinX®-200 at harvest is approximately 175 L. As shown in Figure 2 and Figure 4, cells cultured in the two different bioreactors exhibit very similar growth curves, with three distinct growth phases: an initial rapid growth phase within the first 72 hours, followed by a slower growth phase, and then a plateau phase where cell viability gradually decreases until harvest. The cells are harvested when they have a viability ranging from 93.62% to 93.1%. Both cultures achieved the expected outcomes.

As shown in Figure 3 and Figure 5, both experiments exhibit a similar upward trend in pCO2. During the culture process, the carbon dioxide partial pressure (pCO2) curve is shown in the figure. In the early stages of culture, cells produce lactate to lower pCO2 and maintain pH. Conversely, as lactate concentration decreases and feeding operations increase the concentration of salts and bases, pCO2 will increase.


The results indicate that the CytoLinX® BR 50-2000 single-use bioreactor can be used for both seed culture and large-scale production. The cell culture in the two batches of reactors showed similar cell density, viability, metabolite concentration, and antibody yield as the satellite tank. In two consecutive experiments, the CytoLinX® BR demonstrated stable process control and efficient protein expression, meeting the production expectations of the customers. The CytoLinX® BR series bioreactors exhibit excellent stability in process parameter control for cell culture, providing a reliable and efficient solution for the amplification and production of recombinant proteins and antibodies.

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