Clarity in cell line development

Eliminating enrichment steps in cell line development – reduce cell screening from 5000 to 200 clones per project

Coffee Morning Chat with Ferenc Boldog (ATUM) and Ian Taylor (Solentim): An expert discussion on the evolution of cell line development to negate the need for enrichment

In the time since my previous blog following the ESACT conference in May, in which I discussed fluorescence screening of clones in cell line development, I have been asked several times about the enrichment of cells before single cell cloning. It therefore seemed a beneficial exercise to create a brief video, in collaboration with ATUM, to discuss the current best practice for isolating a population of high-producing clones:

This new paradigm removes the need to screen thousands of clones or to use fluorescence (via FACS, ClonePix or within picodroplets), to detect the highest producers at the single cell level. In fact, many argue that this is a meaningless step at this stage in any event. The most these techniques could reveal at the cloning plate stage would be a crude stratification of high/medium/low productivity normalised to confluence of a colony.

At the recent PEACe Conference in Newport, RI, Dr Balasubramanian, Cell Line Development Scientist, ATUM, illustrated in her presentation the “old-fashioned” philosophy of random integration cloning, highlighting the number of clones that would need to be screened to have a high chance of finding the highest producers. As you can see in Figure 1 below, this approach, when using the Berkeley Lights Beacon® system, would require the sampling of at least 5,000 clones for a 99% probability of isolating the top clone.

In contrast, Dr Balasubramanian demonstrated that by using transposases (non-random integration approach) coupled with the VIPS instrument for single cell isolation, it is possible to directly find the highest producers by screening less than 200 clones per project (i.e. using only 2-3 x 96 well plates) with the added workflow benefit of immediate up-front assurance of clonality.

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