Cell line development

Reducing timelines in generating clonal cell lines for therapeutic protein production

Introduction

In the biopharmaceutical industry, many biological drugs are produced in mammalian cells. Cell line development (CLD) is used to determine which cell lines have the highest recombinant protein production and are also stable during large-scale manufacturing.

In the CLD workflow, cell lines undergo incremental scaling of culture from static to shaker flask expansion and further scaling up in bioreactors. Cell culture in early CLD is currently limited to static format as the size of the well plates and number of cells limit the ability to agitate the culture.

Dynamic High-throuput screening in early stage

Traditional workflow

Traditionally, CLD starts from single cell isolation in post-transfection pools. After clones selection, the outstanding cells are transferred to larger and larger cultivation system, and it almost takes several weeks.

cytena WORKFLOW- C.BIRD™

To improve and accelerate the cell line development, we offer the comprehensive product portfolio for significantly increasing the speed, quality and efficiency of your cell line development workflows.

01

Comparability with
Shaker-flask

CHO-K1 mAb expressing cell line was used for the study. The cell line was adapted to suspension culture in a chemically defined and animal-component-free medium (CD Hybridoma medium #11279-023 by GibCo). Standard 96-well plates (Eppendorf #0030730011, Germany) were used for all experiments.

Comparison studies were performed with cells cultivated in standard static culture, 30 mL shaker-flask culture (shaking speed: 130 rpm; orbit:19 mm) and C.BIRD™ suspension culture in a 37°C, 5% CO2 incubator environment.

Conclusion

The C.BIRD™ enables early transition to suspension cell culture with higher cell growth rate in standard 96-well plates, potentially providing better translation to large-scale shaker-flask/bioreactor conditions for later CLD processes.

02

High-yield protein production

Case 1: The initial cell concentration for both plates (static culture and c.bird TM suspension culture) was 1.5x106 cells/mL. Both plates were placed into an incubator and cultured for 3 days. Cell density was measured by Bio-Rad TC20™ Automated Cell Counter. For protein yield determination, supernatant samples were measured using Abcam Ab10047 ELISA kit and SpectraMax iD3.

Case 2: The initial cell concentration for both plates (static culture and C.BIRD™ suspension culture) was 0.5x106 cells/mL. Both plates were placed in an incubator and cultured for 5 days. Cell density was measured using a hemocytometer. For protein yield determination, supernatant samples were measured with PAIA Biotech PA-104 kit and SYNENTEC ® NyONE.

Conclusion

Compared to traditional static culture, the results show that C.BIRD™ suspension culture enhanced cell growth and significantly increased recombinant protein yield and volume-specific productivity (QP) per cell. Overall, -the system is compact and can easily integrate into established CLD workflows for improved cell line performance.

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