Process Development
Workflow
Cell lines are significant investments. Your newly created cell lines may synthesize a product of interest efficiently, but the therapeutics discovery process is far from over. Commercial production development involves optimizing growth media and feed to a high-producing cell line, harvesting your product of interest, while analyzing the product for quality control. Every step of this process involves repeated cycles of trial and error. The entire workflow must be consistent and reproducible, while maximizing product yield.
Media Formulation and Feed Optimization
Challenge: Identifying the most efficient combination of cell line, growth medium and cell feed.
Proper cell nutrition is essential for cell line stability and high product yields. Cells are taken from previously developed cell lines in a Master Cell Bank (MCB) and inoculated into new media. Optimizing your media parameters is important, knowing that small changes in media composition can have large effects on overhead cost, cell viability and your final product.
As media and feed trials progress, underperforming cell lines from the MCB are removed. Media trials identify both ideal culture conditions and top candidate cell lines. This often requires manual work and can be time consuming, with multiple rounds of human input and trial-and-error before an ideal media composition and cell line combination is found.Once established, cultures producing your product need to be fed. Continuous cultures are constantly changing as cells grow in and metabolize media. Feed composition, timing between feedings and feed volume can change the composition and yields of the product of interest. Correct feed parameters can:
- Maintain cell health and productivity
- Slow buildup of toxic metabolic waste products in culture
- Ensure the product of interest is expressed as you expect.
We can work together to automate this process, using benchtop miniature bioreactors and cell viability analyzers to instantly collect useful data.
Harvest and Purification / Recovery
Challenge: Developing a quick, effective and high-yielding purification procedure.
When appropriate concentrations of your product are present in culture supernatant, cultures are ready for recovery. First, cultures are centrifuged and filtered to remove cells and coarse debris. Supernatant is further purified on a column to capture Protein A, remove viral particles and separate your product from other impurities. Column purification can vary in scope and length; for example, mAbs bound to an affinity purification column may require multiple buffer polishes before elution.
Analytical Development
Challenge: Keeping your product consistent while increasing production scale.
While production is occurring, analyzing your product of interest will save time and ensure product specificity. After your ideal cell growth medium is matched to the ideal cell line, cell culture is dramatically scaled up. Cell lines may not be static in large culture. They may mutate, losing biosynthetic gene clusters or altering product composition and quality.
For example, performing qPCR will ensure the copy number of your gene is consistent as culture volumes increase. Leveraging automation will allow you to quickly characterize quality and function of mAb products in live-cell assays. We can help you develop analytical methods to study your cell lines to ensure consistent quality control alongside production and ease eventual technology transfer.
Summary
- Media and feed must be optimized for your new cell lines. This can be automated and done in parallel.
- Recovery of your product is critical, and often time consuming. Solutions can be created that match your unique product.
- Analyzing cultures while they grow will help you understand your production process. We can help you select or design methods exclusively for your product.
Products and methods described are not intended for use in diagnostic procedures.
