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Cell line development is a cornerstone of modern biotechnology, enabling scientists across academia and industry to create reliable, high-yield cell systems for producing a wide range of biologically derived products.
The production of reagents, such as therapeutic proteins, antibodies, and viral vectors, can be achieved through two primary methods: stable cell line production and transient expression. Each approach has its own set of advantages and drawbacks, which make them suitable for different applications depending on the required scale, speed, cost, and quality of the product.
Stable production cell lines are engineered to continuously express and secrete a product over long periods. In contrast, transient production approaches might be preferred during early-stage development.
| Feature | Stable Cell Line Production | Transient Production |
|---|---|---|
|
Production duration |
Long-term, continuous production (months to years) |
Short-term, transient (days to a week) |
|
Speed |
Slow, requires months to develop a stable line |
Fast, can produce in a week or two |
|
Scalability |
Highly scalable for large-volume production |
Suitable for small to medium-scale production |
|
Consistency/Quality |
High batch-to-batch consistency, stable product quality |
Variable yield and quality between batches |
|
Flexibility |
Less flexible, new cell line needed for different products |
Highly flexible, can produce multiple products rapidly |
|
Cost per Unit |
Lower in the long run, but higher upfront costs |
Higher per unit due to lower yields |
|
Applications |
Large-scale manufacturing (e.g., biopharmaceuticals, antibodies) |
Research, early-stage development, small-scale production (e.g., reagent production) |
After creating an expression plasmid carrying the target protein(s) as well as antibiotic selection markers, the host cells are transfected. Depending on the host cell and characteristics, this can be achieved via different methods, such as nucleofection. Cells not transfected are eliminated from the cell pool by applying antibiotic selection. The transfected plasmid will (semi)-randomly integrate into the genome of the host cells, creating a heterogeneous mix of stably transfected cells.
After establishing a pool of stably transfected cells, it's crucial to isolate single cells and grow them into clonal cultures, since the initial pool contains a mix of cells differing in various traits. These may include growth properties, production level of the target protein(s), as well as quality attributes of the product, among others.
Our automated single-cell cloning systems utilize miniature culture chambers for easy and reliable monoclonality verification, including whole-chamber images and related documentation. All tedious pipetting tasks are automated, streamlining the process and ensuring process consistency.
Established clonal cell lines will undergo various tests to qualify them with regard to growth properties and production of the target product. Different methods can be utilised here, though often antibody-based methods, such as ELISA, are used. The number of tested clones can vary though often several hundreds are analysed to maximise the chance of obtaining high producers that remain stable over time.
Qualifying selected clones need further upscaling in small-scale bioreactors that mimic culture conditions of larger production tanks. The small-scale cultures, often utilizing volumes of 10s to 100s of ml, allow further process development and asses the stability of clonal cultures under more relevant production conditions to further ringfence high-performing clones. Selected lines will undergo further scale-up, or scale-out procedures to increase production capacity.
Want to find out more about our single-cell platforms? Contact one of our knowledgeable experts today, visit our resources page, or request a demo to see the platforms in action.