insect cell expression at low temp

Hi everyone,

I wanted to try to optimize expression of a particular target by lowering the growth temperature of our insect cells.   Does anyone do this routinely?  What temperature do you use?  When do you lower it - at infection or after growth arrest?   I guess the cells grow much slower so does it take longer to reach maximum expression?

 

thanks!

 

peggy

Posted on 29-Mar-2012 8:43 CEST

Hi Peggy,

We use 21oC and 27oC for both Sf9 and Hi5. We lower the temp when we seed and infect for expression. The cells will grow slower which will help for some proteins. We do observe higher solubility or secretion at lower temp for some proteins. We also delay the time of harvest for 24 hrs at 21oC.

Best regards,

Linda

Posted on 18-Apr-2012 6:47 CEST

Hi Peggy and Linda,

I am interested in expressing proteins in insect cells but I would need to go lower than 20C, is that possible? which would be the lower temprature I could work at?

 

Thanks

 

Ariel

Posted on 20-Apr-2012 19:36 CEST

Hi Ariel,

 

We have only tried expressing at 21, and that seems to work well.  Maybe Linda or someone else has tried lower temps?

 

best,

peggy

Posted on 26-Apr-2012 17:35 CEST

Hi Ariel,

Same for us, we have only tried 20-21oC for expression. The cells will grow at temp lower than 20oC but will be very slow. I am interested to hear how you go if you test it out.

Best,

Linda

Posted on 09-May-2012 23:40 CEST
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