Recombinant glycoproteins expression in insect cell....

James Fethiere James.Fethiere at BRI.NRC.CA
Mon Oct 2 23:09:24 EST 1995


Very interesting discussion going on there.  This question really needs some 
clarification. 
In my first message I 
stipulated that my glycoprotein was highly sialated; but did mention,
and probably not stress enough, that these evidences were based solely
on neuraminidase treatment.  Not being a glycoscientist, that's all
I could deduce from the results.  However this is the second glycoprotein
that we express in sf9 cells for which we see addition of sialic acid.  In 
both cases the harvesting was done 72hrs post infection, time at which one
should expect addition of complex sugars according to the study mentioned
above.  So I would like to believe these results are real; what would be
a good reason not to rely on enzymatic treatment or blots to propose the 
presence of these complex sugars in sf9 cells.  Is the neuraminidase from
Boerhinger not pure enough or is it a specificity problem.
I sure would like to get a clearer picture of the situation so I can have
stronger arguments to present to those making the grimace when they see
these results.

Lets here some more comments!

../James


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