Rapid Development and Scale-Up of Biosimilars: A Case Study of Cell Line and Process Development

Increasing availability of genomic and transcriptomic resources for industrially important cell lines coupled with advances in genome editing technology have opened new avenues for cell line development. These developments are poised to help biosimilar manufacturing, which requires targeting pre-defined product quality attributes, e.g., glycoform, to match the innovator's range.

Development of Biosimilar Cell Lines

The high prices of biopharmaceuticals or biologics used in the treatment of many diseases limit the access of patients to these novel therapies. One example is the monoclonal antibody trastuzumab, successfully used for breast cancer treatment. An economic alternative is the generation of biosimilars to these expensive biopharmaceuticals. Since antibody therapies may require large doses over a long period of time, robust platforms and strategies for cell line development are essential for the generation of recombinant cell lines with higher levels of expression.

Production cell line development starts with expression vector construction and transfection. After being transfected with plasmids bearing the recombinant genes, as well as selection marker(s), cells are screened for growth and productivity following growth recovery, serum-free suspension adaptation, amplification (if necessary), and clonal selection. The screening and selection of a highly productive and stable clone from the transfectant population in a limited time frame is a major challenge.

Rapid Development and Scale-Up of Biosimilars: A Case Study of Cell Line and Process Development Fig. 1 Illustration of a typical process to develop a mammalian cell line for recombinant protein manufacturing.

At Creative Biogene, multiple biosimilar stable cell lines have been developed and have shown high performance in yield. In addition, more biosimilar stable cell lines are under development.

Cat. No.	Product Name
CSC-BS0001	Rituximab Stable Cell Line - CHO-K1
CSC-BS0002	Trastuzumab Stable Cell Line - CHO-K1
CSC-BS0003	Cetuximab Stable Cell Line - CHO-K1
CSC-BS0004	Nivolumab Stable Cell Line - CHO-K1
CSC-BS0005	Pembrolizumab Stable Cell Line - CHO-K1
CSC-BS0006	Adalimumab Stable Cell Line - CHO-K1
CSC-BS0007	Infliximab Stable Cell Line - CHO-K1

The Benefits of Biosimilar Stable Cell Lines

Biosimilar cell lines have several advantages in the production of biosimilar drugs.

Firstly, they offer increased efficiency and cost-effectiveness compared to traditional methods. By utilizing well-established and optimized cell lines, it becomes possible to streamline the drug development process and reduce development time and costs.
Additionally, biosimilar cell lines enable reproducibility and consistency in drug production. The use of a well-defined cell line ensures uniformity in terms of protein expression, glycosylation patterns, and post-translational modifications. This allows for greater predictability in the drug manufacturing process, resulting in a reliable and consistent product.
Moreover, biosimilar cell lines provide flexibility in drug design and development. These cell lines can undergo genetic modifications, such as gene knockouts or overexpression, to enhance productivity, optimize protein quality, or modify specific attributes of the biosimilar drug. This flexibility enables customization of the biosimilar drug to meet specific requirements and potentially improve clinical outcomes.

Process Development Steps for Biosimilars

Step 1: cell line selection and engineering

One of the most crucial process development decisions made during the development of a biosimilar is the choice of cell line. The cell line is one of the key determinants of the glycosylation patterns of biologicals, making the choice of the mammalian expression host very important in determining the final glycoform profile of the biosimilar. For example, Chinese hamster ovary-based cell lines are the most popular for the development of biological drugs because they generally produce similar glycosylation patterns to humans and have several advantages, including their ability to grow in suspension, their high specific yield and their stability to changes in pH and oxygen.

Step 2: cell culture process development

The cell culture process involves thawing a vial of frozen cells from a cell bank. The vial is inoculated into shake flasks to increase the cell density, and the cells are then grown in serial sub-cultivations until the target production scale is reached. The cells are maintained in growth media and are provided with the required nutrients and additives to ensure the viability of the cells.

Step 3: purification process optimization to guide CQAs into similarity range

Once the cell culture phase is completed, a purification process is used to recover the target protein, while removing unwanted impurities, including adventitious viruses, host cell proteins and DNA, aggregates and endotoxins.

Step 4: achieving a stable formulation

After the product is purified, it is concentrated and formulated using ultrafiltration and diafiltration.

References

Lao-Gonzalez, T.; et al. Screening and selection strategy for the establishment of biosimilar to trastuzumab-expressing CHO-K1 cell lines. AMB Express. 2021 Jan 3;11(1):1.
Gotham D. Cell line access to revolutionize the biosimilars market. F1000Res. 2018 May 3;7: 537.
Vulto, A. G.; et al. The process defines the product: what really matters in biosimilar design and production? Rheumatology (Oxford). 2017 Aug 1;56(suppl_4): iv14-iv29.
Lai, T.; et al. Advances in Mammalian cell line development technologies for recombinant protein production. Pharmaceuticals (Basel). 2013 Apr 26;6(5):579-603.

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

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Rapid Development and Scale-Up of Biosimilars: A Case Study of Cell Line and Process Development

Development of Biosimilar Cell Lines

The Benefits of Biosimilar Stable Cell Lines

Process Development Steps for Biosimilars

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