We had experience with a relatively small glycoprotein - when glycosylation sites were deleted, solubility went drastically down - we could not express soluble any more. Back to eukaryotic expression system which worked.
So may be you were really lucky. Jan On Tue, Apr 11, 2017 at 10:34 PM, Bernhard Rupp <hofkristall...@gmail.com> wrote: > Hi Fellows, > > > > a humble question for our glyco-expressionists: > > > > I have mutated out the Asns of the N-glycoslation consensus sites for Asp > > (Asp simply because the PNGaseF treated protein stays stable so I thought > that might be a good guess) > > and indeed the unglycosilated mutant expresses well and gets secreted as > planned. > > > > But rumor has it that glycoproteins that are mutated to non-glyc often are > not processed correctly and > > that we had just dumb luck. > > > > May I poll the educated opinion of the erudite here? > > > > Cheers, BR > > > > ------------------------------------------------------ > > Bernhard Rupp > > Crystallographiae Vindicis Militum Ordo > > http://www.hofkristallamt.org/ > > b...@hofkristallamt.org > > +1 925 209 7429 <(925)%20209-7429> > > +43 767 571 0536 <+43%207675%20710536> > > ------------------------------------------------------ > > :(){ :|: & };: > > ------------------------------------------------------ > > > -- Jan Dohnalek, Ph.D Institute of Biotechnology Academy of Sciences of the Czech Republic Biocev Prumyslova 595 252 50 Vestec near Prague Czech Republic Tel. +420 325 873 758
