Protein production: scale-up

Duncan Clark Duncan at genesys.demon.co.uk
Mon Jun 12 10:24:46 EST 1995


In article: <1995Jun11.105322.8890 at mbcf>  heath at mbcf.stjude.org (Richard_Heath) writes:


> If I remember from my PhD days, the switch from shake flask to fermentor
> changes things a lot! (Even swithching from one size shake flask to another
> alters things - mainly due to aeration efficiency, I think) However, scaling up
> fermentors should not be too much of a problem...  Thus, I would forget the 
> shake flasks pretty much altogether, and optimise in a small fermentor, then 
> go up to the big'un (I only got as big as 30 liters, and optimised in a 2 
> liter).  Things like the medium will probably remain the same - aeration is 
> going to be one of the biggest factors.  You can, of course, also maintain a
> stable pH/aeration etc in the fermentor.

It may sound easy ie 2 litres to 20l etc but going up further will be more 
difficult ie change in baffle size, impeller design/size etc etc. I would 
monitor everything, OD, dO2, dCO2, pH, rpm, temp etc If possible avoid using
an expression system that needs Ampicillin. Go for kanamycin/neomycin. Amp
is destroyed in minutes so you may land up with just E.coli minus your 
expression plasmid! Definately avoid shake flasks.

Good luck
Duncan 

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