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Antibody production in Expi293 cells


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#1 tom43

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Posted 02 September 2013 - 11:53 AM

Hi,

 

we try to produce antibodies in the new Expi293 suspension cells (Life Technologies) by transient transfection with two pFUSE expression vectors for light and heavy chain. The cells suffer from high cell death with only ca. 65% viability 3 days after addition of DNA:ExpiFectamine complex. There are also dead cells by only adding the ExpiFectamine. Is there anyone else with comparable observations? Any idea for optimizing the experiment to reduce cell death?

 

Many thanks!



#2 bob1

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Posted 02 September 2013 - 12:36 PM

Yeah, the "*fectamine" range of reagents seem to be quite toxic generally, especially Lipofectamine  and Lipofectamine 2000.  The exception so far is lipofectamine RNAiMax.  You could try a different transfection reagent such as Fugene (Promega), PEI (Sigma, and a number of other companies), or any of the electroporation systems.



#3 tom43

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Posted 06 September 2013 - 01:08 PM

Hi Bob, thanks for your reply. Yes, we also made the observation with the Lipofectamine 2000. Hi transfection rates, but toxic. It should be somehow different with the Expi293 Expression System, which is based on HEK293 cells selected for high-density growth (= "Expi293") and a specifically adapted transfection reagent called ExpiFectamine293 (plus so-called enhancers). This "kit" was arranged by the manufacturer so I wonder about the high cell death rate in our lab...

 

As a next step we will have a look at the ExpiFectamine293 compared to HEK293 transfection reagents from Mirus, PolyPlus etc.. I will post our results once finalized.

 

In parallel we tested Freestyle MAX CHO-S cells, but these cells seem to produce much lower antibody using transient transfection. Nevertheless we don´t observe any toxicity during transfection in the CHO cell experiments.






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