What is it about?
Manufacturing cell lines are the single most important component of every biologics manufacturing process. These cell lines should express the protein drugs or vaccines at high and consistent level. The paper describes a cell line development process, based on Leap-In transposases, that consistently delivers genetically stable high producer cell lines within a short timeframe. We also demonstrate the comparability between the intermediate stable pools and the derivative clones. This allows initiating PD work, formulation development, toxicology material or even Phase 1 clinical drug substance manufacturing using representative cell substrate while clone generation is still in progress.
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Why is it important?
The main advantages of the system are: - multiple, but always single copy integration sites into transcriptionally active open chromatin regions - robust genetic stability - no payload limit. Multi-ORF expression constructs enable to control protein subunit ratios that are consistent between all the recombinant cells and the original stable pools. With other words, every integration site in every recombinant cell within a pool expresses the subunits at the same ratio. It is particularly important for the novel multi-specific protein drug architectures. -as a consequence of clonal homogeneity, the parental stable pools produce highly comparable material enabling early CMC activities, and material generation (tox or even Phase 1) - the development timelines are among the shortest compared to industry standard. The transfected pools recover significantly faster, and screening ~50-100 clones is sufficient to identify high producer stable clones. This is in stark contrast to the industry standard where thousands of clones need to tested and ranked in order to isolate the rare acceptable clones.
Perspectives
We believe that this development process can change the current manufacturing cell line development paradigm. - Critical decisions are made during stable pool ranking. Since the pools and the derivative clones are highly comparable once a pool with the desired features CQAs is found, clone isolation and subsequent clone ranking will match the characteristics of the parental pool. This allows testing multiple pools in case of complex molecular architectures and selecting the best one within a short period of time. Once it is done, clone isolation will deliver the desired clones. - This, pool to clone, comparability also allows to utilize the stable pools for process and formulation development. - Using the stable pools for material generation. The comparability of stable pool and clone derived drug substance enable to manufacture representative drug substance using pools. Construct synthesis and pool generation is about 4 weeks to allow fast manufacturing. This feature is especially important under emergency circumstances and Leap-In mediated pools have already been utilized in quick Covid 19 response related programs.
Ferenc Boldog
ATUM
Read the Original
This page is a summary of: Accelerating and de‐risking CMC development with transposon‐derived manufacturing cell lines, Biotechnology and Bioengineering, April 2021, Wiley,
DOI: 10.1002/bit.27742.
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