Advanced Flow Cytometry: Your Fast-Pass to Stable, High-Producer Cells
The global market for biologics, such as monoclonal antibodies and antibody-drug conjugates, is growing. However, developing and manufacturing these essential drugs is expensive, and the risk of failure can be high. In this blog post, we discuss common cell line development challenges and explain how advanced flow cytometry can accelerate the drug discovery process with the potential to bring high-quality biologics to patients sooner.
This article is posted on our Science Snippets Blog
What are the steps in a cell line development process?
Cell line development is the cornerstone for producing therapeutic proteins, antibodies, and other biomolecules. The workflow involves multiple steps:
- The development process begins with cloning genes for the desired bioproduct into an expression vector, and then transfecting it into a mammalian host cell line. Chinese hamster ovary (CHO) cells are widely used for biologics production because they are relatively easy to culture, give high yields, and can replicate the post-translational modifications and folding found in human proteins.
- Stably transfected single cells are isolated for clonal expansion and assessed for parameters like antibody titer, viability, and growth.
- After identifying the high-productivity clones with optimal critical quality attributes (CQAs) and target specificities, it’s time to optimize the culture conditions for peak cell growth and protein production.
- Next, one or more of the best-performing clones are characterized for stability, productivity, and key properties. For example, antibody binding kinetics are often measured using protein analysis techniques like BLI or SPR, which are a fast way to get more biologically-relevant data.
- Finally, the cells are banked in accordance with good manufacturing practices (GMP) ahead of scale-up and commercial manufacturing.
What are the common challenges in cell line development?
The first challenge is choosing a cell line that can create all the desired features of the biologic. These include cell stability, growth characteristics, media requirements, and whether the cell line has a documented history.
Identifying the best-performing clones is another difficult step. One way to increase the likelihood of discovering rare, highly productive cells is by screening large numbers of clones. But more samples mean longer timelines for process development, especially when samples must be sent away to an external provider for analysis.
The next hurdle is characterization. Comprehensive bioanalysis is crucial to making sure the final product is safe and effective, but it often involves repeated sampling, running assays across multiple instruments, and integrating all the datasets for analysis. This uses up precious samples, takes time, and in the end, may not accurately predict outcomes during scaleup.
How does advanced flow cytometry address process bottlenecks?
A traditional cell line development workflow might involve low-throughput immunophenotyping of cells with a standard flow cytometer, evaluating viability using a plate-based assay, and measuring antibody titers by ELISA. Performing these assays and piecing together the disjointed datasets could take days or even weeks.
In contrast, advanced flow cytometry systems combine multiple readouts in the same well of a multi-well plate, providing insights on both phenotype and function from a single experiment. Not only does this reduce sample and reagent requirements, but it also gives researchers a broader overview of each clone in less time.
iQue® Advanced Flow Cytometry platform
The iQue® Advanced Flow Cytometer is a next-generation high-throughput flow cytometry platform that addresses bottlenecks and turns up the speed dial on the cell line development process. With a capacity to analyze up to 15 parameters per clone, using as little as 5 μL of sample (in a 384-well plate), it can give you both phenotypic and functional information in one experiment.
What really sets the iQue® instruments apart is the fast sample processing: 5 minutes for a 96-well, and 20 minutes for a 384-well plate in 20 minutes. The instrument is also really easy to use, from setting up the experiment all the way to data analysis. The integrated software simplifies the entire data analysis workflow, so you can visualize data across the entire plate and easily adjust gates in real time.
Your fast-track lane to the top-producer cells is up ahead
The rising demand for novel biotherapeutics highlights the need for solutions that streamline the cell line development workflow. Technologies like the iQue® platform transform timelines by taking you from sample, to raw data, to insights with exceptional speed and ease.
Learn more about the iQue® platform as well as multiplex products like the iQue® Cell Count and Viability Kit and the iQue® Human IgG Titer and Viability Kit, that can help you quickly evaluate clones.
