IUBio

optimization for recombinant protein expression

Ian Mc i.mcfarlane at icrf.icnet.uk
Sat Mar 20 10:07:47 EST 1999


Hi Robert,

Can you post some more information on this DOE?

I am interested,

Ian Mc


In article <slrn7erlu0.3m4.rbb at localhost.localdomain>,
rbb at nebiometrics.com wrote:

> On 14 Mar 1999 04:01:21 -0800, khlee at MED.CHUNGBUK.AC.KR <khlee at MED.CHUNGBUK.
> AC.KR> wrote:
> >Dear Netters:
> >
> ><P>I am trying to express a recombinant protein on pATH vector (TrpE
promoter)
> >in E. coli.
> ><BR>This protein will be induced in M9 media, which contains casamino acid
> >in 0.5% (5g/L) as usual concentration.
> >
> ><P>(1) I wonder if I can increase the yield of recombinant protein when
> >I increase the concentration of casamino acid in M9 media.
> >
> ><P>(2) In addition to the minimal salt, I usually add glucose, thiamine,
> >calcium and magnesium. Is it required to increase the concentration of
> >salt and other additives if the concentration of casamino acid shall be
> >increased?
> 
> The answer to your questions can be determined experimentally using design
> of experiments (DOE). DOE is a method for systematically varying the
> system parameters (i.e. medium composition) so as to find the optimum
> output (i.e. yield of recombinant protein).
> 
> The advantages to using DOE are that it is faster and easier than
> traditional methods and you generally obtain more useful information. If
> you are interested, let me know and I will send more information.
> 
> -- 
> Robert Burrows
> rbb at nebiometrics.com



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