Sharing experience

on recombinant protein development challenges

Is my hybridoma the ideal system for production of my therapeutic MAb?

Published on 2016-02-16 Upstream Process

Is my hybridoma the ideal system for production of my therapeutic MAb?

What you will read about: our opinion on the best system to use for production of your MAb candidates for POC studies.

When after an intense R&D effort, you have a high potential therapeutic MAb, obtained from a good-producer hybridoma, all you want to do next is a conclusive Proof-Of-Concept that will convince investors.
That POC should bring credibility to your idea. It will probably be short and will not necessarily be thorough due to time and money constraints.  

In that context, it may be tempting to call upon your heard-to-earn hybridoma, since only limited quantities of MAb are necessary. By experience such a choice, dictated by IP, budget and time constraints may be far from the best.

Indeed the hybridoma will quickly produce the 10 to 100 mg you need for your demonstration. But given what is at stake it may be worthwhile to consider the risks associated with the use of hybridoma for such developments.

What are some of them?

Nowadays, no one wishes to develop a therapeutic MAb by hybridoma culture. To our knowledge, only RemovabTM  is still produced by it’s original hybridoma. All others have been re-engineered into recombinant.
In fact hybridoma production no longer addresses several of the current concerns about Mab development, which imply the parallel evaluation of variants: fusion approaches, isotypes, humanization variants, …)

Morever, from a hybridoma production, it is impossible to predict the final product characteristics such as proteolytic degradation, glycosylation profile, aggregation… all of which, by experience will impact not only the purification process (to remove the aggregates, impurities, …) but also the in vitro or in vivo activities.
So in therapeutic development, the hybridoma route may be dangerous to follow too long because of these specific hurdles. Clearly, hybridoma is not a good model for therapeutic development and the MAb produced for the preclinical phase may turn out to be quite different from those obtained during the POC, which is then wasted.


If you want a POC that is representative of the product to be developed, forget the hybridoma and turn to something that will look like and feel like the final product, which therefore should be produced by the expression system in use in the industry.
Transitory transfection is quite suitable at this stage of parallel testing, fast and with good yields. The only limitation is when the demand exceeds 100mg: production costs increase rapidly after that scale due to the cost of the DNA preparation and of the transfection agent.

At GTP we have chosen and would recommend the CHO Xpress technology. We call it « transi-stable » because it combines the advantages and transient and stable transfection. Indeed with this technique super-producing stable clones are generated without cloning, so very rapidly. The obtained clone can be stored and re-used later for subsequent or larger needs, with an immediate and significant cost benefit compared to transient transfection, starting from relatively small scale.

The technology finds a sweet spot of entry during a POC, since it will result in a molecule close to the final biotherapeutic, in as short as 3-4 months, and offers the capacity to economically produce all that will be needed during the preclinical development (up to gram scale).

If you take our advice for your next POC, you will put aside your hybridoma and move on with transitory transfection in CHO cells or better still, with CHO-Xpress!