Cell line development: pitfalls, challenges and solutions
Cell line development plays a key role in the identification of biological targets and commercial manufacturing of biotherapeutics. Despite its importance, the process faces a number of challenges, particularly when it comes to achieving monoclonality and assessing productivity.
In this interview, we spoke with Darius Wilson, Product Manager of CellCelector Systems at Sartorius (Göttingen, Germany), to explore how technological advances, like the CellCelector CLD from Sartorius, are helping to overcome these issues to streamline and de-risk monoclonal cell line development.
Darius Wilson
Product Manager, Product Management BioAnalytics Workflow
Sartorius
Specializing in early identification of disease progression resulting from a dysregulated host response to localized bacterial or viral infections, Wilson is the author of 14 patents and numerous publications in fields such as gastrointestinal surgery, trauma-related complications, antibiotic guidance and emergency and intensive care medicine.
He obtained his Ph.D. in physiology and pharmacology and an MBA from Glasgow Caledonian University (Scotland).
- Why is monoclonality so important in cell line development?
- How do researchers currently identify monoclonality, and what challenges do they face in doing so?
- How is productivity usually assessed in the cell line development workflow, and what issues can arise at this stage?
- Are there any other significant bottlenecks in the cell line development process?
- How can cell selection technologies like CellCelector CLD help to overcome these challenges?
- Do you have any advice on how to get the best results from CellCelector CLD?
Why is monoclonality so important in cell line development?
Monoclonality is fundamental within cell line development, as it ensures uniform protein expression levels, consistent post-translational activity and stability, and results in cell lines being less prone to genetic instability or mutations that can occur during long-term culture. In short, ensuring cell lines are derived from an initial single cell ensures that the final biologic is not contaminated with any unwanted cell populations or adventitious agents, thereby ensuring overall safety and preventing potential contamination, which could lead to a less effective biological product.
How do researchers currently identify monoclonality, and what challenges do they face in doing so?
There are several methods used to typically identify and isolate single cells. Traditional methods include limiting dilution and fluorescence-activated cell sorting (FACS), which can be both tedious and time-consuming, require extensive sample preparation and often result in poor cell viability and proliferation. More automated solutions can significantly decrease the time required to generate a monoclonal cell population and monitor subsequent growth from a single cell to a highly productive clone.
How is productivity usually assessed in the cell line development workflow, and what issues can arise at this stage?
Productivity in cell line development is assessed by measuring the amount of target protein generated by a specific cell line, against time, volume or cell number. This is generally assessed further upstream by high-throughput methods such as Enzyme-Linked Immunosorbent Assay (ELISA) or Protein A HPLC. The disadvantage of this process is that no prior pre-selection is conducted, resulting in the measurement of hundreds or thousands of clones, which can be extremely time-consuming.
Are there any other significant bottlenecks in the cell line development process?
Yes, absolutely. Other significant bottlenecks include, and are not limited to:
- Clone productivity – the vast majority of clones may initially have a low productivity, which makes early clone selection problematic.
- Loss in clone productivity – after identifying initially high-producing clones, clones may lose productivity over time or across passages, or perform well in small scale but poorly when upscaled for manufacturing purposes.
- This therefore results in a high risk of researchers choosing suboptimal candidate clones, which can directly impact project timelines and time-to-market.
In addition, further bottlenecks may include product stability testing, to ensure high productivity and quality is maintained over time, and ensuring that all processes are fully documented to fulfil stringent regulatory requirements.
How can cell selection technologies like CellCelector CLD help to overcome these challenges?
The CellCelector CLD instrument helps to collapse the overall cell line development workflow, by providing users with:
- A visually confirmed monoclonality screen of up to 85,000 single cells within a single plate.
- Excellent clone outgrowth that can then be tracked over 4 days to identify the fastest growing clones.
- An initial clone productivity assessment using an EMA 410 compliant productivity assay, which helps identify the optimal clones early in the process, and thereby decreases the overall workflow duration.
- Up to 100% transfer efficiency and viability of selected clones for further characterization.
Do you have any advice on how to get the best results from CellCelector CLD?
Assessment of monoclonality and initial clone productivity in the earliest stages possible is key to identifying optimal clones and reducing the number of clones that are taken forward for further assessment. Further characterization with the Octet® Platform helps researchers confirm initial productivity assessment on the CellCelector, so that the top candidate clones are quickly identified and are progressed.
This interview is part of a Spotlight on cell line development. Visit the full feature and discover expert insights on the topic.
The interviewee has no competing interests to report.
The opinions expressed in this interview are those of the interviewee and do not necessarily reflect the views of BioTechniques or Taylor & Francis Group.
This content was created in partnership with Sartorius.
