Cell Line Development (CLD) 2.0

6th July 2017

Category: Cell Line Development

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By: Caroline Hauser - Team Leader, Dr Christoph Zehe - Head of Technology Development, Juliana Schubert - Scientist, Kristin Thiele - Scientist,

The production of biologics in Chinese Hamster Ovary cells (CHO) is becoming more and more challenging due to an increasing number of complex and difficult-to-express (DTE) molecules. Nevertheless, general Cell Line Development (CLD) platform processes should provide reliable high-titer cell lines irrespective of the therapeutic protein formats to be expressed. In the last 20 years the growing number of innovator biologics and biosimilars have formed a competitive environment, where speed and efficiency of generating robust and highly productive cell lines need to be improved continuously.

Reducing CHO Cell Line Development Timelines

As mentioned above, a fast and efficient cell line development service is highly sought after by the biopharmaceutical industry. Providing speed whilst still producing a robust, high-producing and stable cell line is a key industry challenge for all those involved in cell line development. Nevertheless, it is achievable and at Sartorius Stedim Cellca, we continuously seek to improve our platform to meet the market demands.

The industry standard timeline for cell line development projects can be anywhere from 6 to 9 months. Another challenge is producing difficult-to-express (DTE) proteins in CHO cells and can lead to delays or in the worst case scenario termination of cell line development.

In an ongoing continuous improvement of our CHO DG44/DHFR platform technology, we have developed an improved cell line development approach capable of reducing timelines to just 5 months for both easy and difficult-to-express (DTE) molecules.

Standard approach vs Cell Line Development (CLD) 2.0

We managed to reduce the timeline compared to standard cell line development by developing a new and robust pool generation approach, as demonstrated in Figure 1. In pool generation stage of the standard approach cells were cultivated using selective medium or medium containing additional 2.5 nM methotrexate (MTX) for three weeks followed by an amplification step for a further 3 weeks.

In our newly developed CLD 2.0 approach we were able to omit the 30 nM MTX amplification step by increasing MTX concentration during the first selection phase of 3 weeks. Removing the amplification step saves us 4 weeks in the pool generation process with no impact on stability or product titer.

Figure 1: Standard approach and new CLD 2.0 process for Cell Line Development at Sartorius Stedim Cellca GmbH.

Download our scientific poster to view the results of our new CLD 2.0 approach.

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