Proof of clonality and its importance in stable cell line development
Clonality is a crucial step in stable cell line development (CLD) for biotherapeutic workflows and one closely monitored by the medicines’ regulators. Their questions lead to two possible conclusions about how they view the importance of clonality1:
- Assurance of “clonality” of stable production cell lines is of major importance in assessing the safety and efficacy of the product1.
- Without adequate proof of “clonality” additional studies of the cell line and product are often required to ensure the product’s quality and homogeneity1. These additional studies and control strategy will also have to be demonstrated for legacy cell lines.
If clonality is not sufficiently evidenced, regulatory bodies such as the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) will require additional manufacturing controls, increasing the cost of clinical trials and delaying drugs from reaching patients. With increasing pressure to speed up drug development and improve drug safety, this can be an expensive and even fatal setback for a company.
Single cell cloning is key in a variety of workflows, not just biotherapeutics and monoclonal antibody production. Companies are increasingly in need of “stable producer” cell lines for gene therapy vector production. As these cell lines will produce a therapeutic (i.e. the virus vector) there is an implied requirement that the cell line must be shown to be clonal for regulatory approval. Similarly, in gene editing workflows involving iPSC (induced pluripotent stem cells) and cell therapies, single cell isolation techniques are important as the therapeutic cells are intended for patient use.
Securing the correct single cell cloning method and instrumentation is clearly essential, but what instruments can be used to streamline this process and achieved a more automated single cell cloning approach?
Our highly innovative instruments have raised the bar for single cell dispensers, providing documented proof of clonality and offering compatibility with a wide variety of single cell cloning protocols. Our portfolio of products (VIPS™ single cell dispenser, Cell Metric® and Cell Metric® CLD cell imaging systems) enable customers to establish a simple to use and completely integrated process: seeding single cells into wells, whole well imaging, production of a clonality report, optimising clonal outgrowth and early monitoring of clone quality attributes.
Our technology removes the need to screen thousands of clones or to use fluorescence (via FACS, ClonePix or within encapsulated picodroplets) to detect the highest producers at the single cell level. Using traditional random integration methods of clone screening at least 5,000 clones would need to be sampled to achieve a 99% probability of isolating the top clone. In contrast, using transposases (non-random integration approach) or landing pads (targeted gene integration) coupled with the VIPS instrument for single cell isolation, it is possible to directly find the top clones (for cell line protein production and stability) by screening less than 200 clones per project with the benefit of immediate and up-front proof of clonality.
This can have a dramatic impact on workflow productivity, as demonstrated by our customer at Janssen R&D. Janssen’s Cell Line Development Group doubled the speed of its workflow using the VIPS, shortening a two-step process to develop cell cultures to a single step, halving development times while still satisfying the clonality requirements of the regulators. Download the full case study here.
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