Dirk Lütkemeyer: Platform Technologies

We are using platform technologies mainly for the purification of recombinant antibodies from mammalian cell culture. In developing the initial platform, we conducted a research project. Up to now we evaluated several different antibodies from CHO or hybridoma cell cultures. The biggest difference we saw was besides the characteristics of the different subclasses, the impurity profile which depended on the specific cell culture conditions that were being used.

With use of platform technologies, some conditions used in the process are standardized while others need to be customized. The conditions that are customized typically depend what is going on with the molecule in terms of the impurity profile, host cell protein, DNA, and aggregates. Steps are adjusted to minimize aggregation of the antibodies, for example, or to get better DNA or host cell protein removal.

In terms of adjusting conditions, we usually first focus on the elution profile. If that doesn’t work, we explore other buffer systems and then if necessary, try different chromatography media. We do evolve the platform. For example, if new chromatography media are available, we look to add to them to the platform.

We find that using platform technologies provides for a faster process development. The system also provides us with insights as to what are the crucial steps in the process. Sometimes a small change in just the buffer conditions has a big impact. The one caution when using platform technologies is to avoid forcing your molecule into the existing platform. It may be the case that better media or methods are available for a particular molecule, so avoid be locked in but rather find the best balance between use of the platform and new approaches when needed.

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1 Comment for this entry

Thomas PONCET
March 14th, 2013 on 5:02 am

Definitely, purification platforms are intended to save time (and money !) by avoiding unecessary optimization time during the first development steps of a new protein (and mainly mAbs to be honnest).
Still, it appears that it is difficult to bring to life a uinque template, as each mAb will exhibit different aggregates and HCPs content, which may force to apply a “too powerful” template to cases where the impurities (post protein A step) are alrready low.
We thus see templates using 3 steps (protein A ==> CEX in bind / elute ==> AEX in flowthrough) for mAbs containing higher levels of HCPs and aggregates, and 2 steps (protein A ==> AEX in flowthrough) when the lucky developer has a low aggregating and already very purified mAb.

And it is also not illogical to have a 3 steps template (protein A ==> CEX in bind / elute ==> AEX in flowthrough) with alternative between two CEX media.
One offering very good aggregates removal, and one showing less selectivity on this specific goal.

As always with chromatography, the solution needs to be customized, as the protein is the one that decides !
The good news being that newly launched CEX media are able to provide the needed aggregates removal, while keeping laoding capacity and yield at the maximum.
This is probably one way to further simplify the templates, and achieve the expected goals.

Good and successful developments to all,
Thomas

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