High-performance CHO Stable Cell Line Development

Chinese Hamster Ovary (CHO) cells have been widely used in the production of biologics. Several features render CHO cells a powerful bioprocess tool such as long-standing safety record, easy to grow in large scale cell culture under optimized conditions, efficient selection system, high productivity, post-translational modifications etc. However, stable cell line generation is a time and cost-consuming process, especially to obtain a single cell-based clone with high and consistent expression level.

Creative Biogene has more than 10 years of experience in stable cell line development and has developed our own proprietary High-performance CHO Stable Cell Line Development (HP-CHO SCLD) platform. Our HP-CHO SCLD platform can be used for a broad range of applications, from R&D development to cGMP commercial production. Based on years of experience and innovation, our scientists are able to customize each cell line development process to meet various client needs and specific requirements. Whether you work in drug discovery, or upstream, or downstream bioprocessing, Creative Biogene can always help you to generate the fit-for-project stable cell lines with high quality and accelerated timeline.

Highlights

Robust CHO-K1 Host Cell Line

Grow in suspension, allowing for large scale culture
Compatible with chemically defined commercial media, animal product-free culturing
Path to commercialization

Proprietary Expression Vectors

Optimized for CHO delivery and expression
Monocistronic, bicistronic vectors

Cell Line Development

Expressing diverse biologics with high-quality, mAbs, biosimilars, recombinant proteins, growth factors, bispecifics, etc.
Great performance in productivity, e.g., 6-8 g/L mAbs
Excellent cell line stability, over 30 generations

Fast Timelines

Generally 12-16 weeks, from DNA to high-expressing stable clones

After the RCB is developed, Creative Biogene also offers cell line characterization services, cell bank generation, and process development services:

Support for assay development and analytics
Development of upstream and downstream processes for scalable production and tech transfer
Cell Bank Generation: PCB, MCB, WCB
Traceable records from vector construction to cell line development which support clinical trials and market authorization (IND etc.)

Service Workflow

Case Study: State-of-the-art Facilities

State-of-the-art Facilities Figure 1. Left): 4D-Nucleofector® X Unit from Lonza for efficient transfecting various types of cells; Right): Flow cytometry analysis of GFP expression in CHO-K1 cells 48 h post-transfection.

State-of-the-art Facilities Figure 2. Left): CloneSelect® Imager from Molecular Devices for high-throughput automated imaging and analyzing cells with accurate, objective and consistent results; Right): Tracking the formation of a colony from a single cell.

Case Study: Cell Line Development & Productivity

ID	Targets	Culture	Titer (g/L)
1	Human IgG1 Kappa	200mL FedBatch (without optimization)	6.41
2	Human IgG1 Kappa	200mL FedBatch (without optimization)	2.6
3	Human IgG1 Kappa	200mL FedBatch (without optimization)	3.77
4	Human IgG1 Kappa	200mL FedBatch (without optimization)	1.8
5	Human IgG1 Kappa	200mL FedBatch (without optimization)	3.86
6	Human IgG1 Lambda	200mL FedBatch (without optimization)	4.08
7	Human IgG1 Kappa	200mL FedBatch (without optimization)	3.63
8	Human IgG1 kappa	200mL FedBatch (without optimization)	4.79
9	Human IgG1 Kappa	200mL FedBatch (without optimization)	5.09
10	Human IgG1 Kappa	200mL FedBatch (without optimization)	3.56
11	Human IgG1 Kappa	200mL FedBatch (without optimization)	5.27
12	Human IgG1 Kappa	200mL FedBatch (without optimization)	0.96
13	Human IgG1 Kappa	200mL FedBatch (without optimization)	2.36
14	Human IgG1 Kappa	200mL FedBatch (without optimization)	8.32
15	Recombinant Protein	200mL FedBatch (without optimization)	1.23
16	Recombinant Protein	200mL FedBatch (without optimization)	0.56
17	Recombinant Protein	200mL FedBatch (without optimization)	0.31
18	Recombinant Protein	200mL FedBatch (without optimization)	0.47
19	Recombinant Protein	200mL FedBatch (without optimization)	0.88
20	Recombinant Protein	200mL FedBatch (without optimization)	6.2
21	Recombinant Protein	200mL FedBatch (without optimization)	0.64
22	Recombinant Protein	200mL FedBatch (without optimization)	0.96
23	Recombinant Protein	200mL FedBatch (without optimization)	3.64
24	Recombinant Protein	200mL FedBatch (without optimization)	2.13
25	Recombinant Protein	200mL FedBatch (without optimization)	0.63
26	Recombinant Protein	200mL FedBatch (without optimization)	0.77
27	Recombinant Protein	200mL FedBatch (without optimization)	4.39
28	Recombinant Protein	200mL FedBatch (without optimization)	1.52
29	Recombinant Protein	200mL FedBatch (without optimization)	0.74
30	Recombinant Protein	200mL FedBatch (without optimization)	0.38
31	Recombinant Protein	200mL FedBatch (without optimization)	1.36

Figure 3. CHO stable cell lines show excellent productivity

Case Study: Cell Line Evaluation

Creative Biogene offers multiple analytical methods to evaluate and select best single cell clones with high productivity and stability. These analytical methods include Western Blot, PAGE, ELISA, HPLC etc.

Cell Line Evaluation Figure 4. Determination of antibody affinity by ELASA. Two batches of antibodies produced from CHO stable cell lines show high affinity with the antigen.

Figure 5. Determination of antibody purity by PAGE. Two batches of antibodies produced from CHO stable cell lines show high purity.

Figure 6. Determination of antibody purity by HPLC. Two batches of antibodies produced from CHO stable cell lines show high purity with 99% and 95% respectively.

* For research use only. Not intended for any clinical use.

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