glycosylation in insect cells ?

Gregory P. Adams, Ph.D. gp_adams at fccc.edu
Tue Jun 29 02:57:02 EST 1999


Hi,

    I'm sorry to tell you this, but insect cells do glycosylate proteins
differently than mammalian cells (and do alter the epitopes accordingly).
Furthermore, the type of insect cell employed will also alter the
glycosylation.  We had similar problems trying to express HER2/neu ECD in
sf9 insect cells.  We have finally gone over to using mammalian expression
systems.  But you also have to be careful there as some cell lines and
promotors don't work well for some proteins.

Good luck!

-Greg Adams

Svend Kjaer wrote:

> Dear all,
>
> I am expressing the extracellular portion of a human receptor in
> baculovirus-infected insect cells as an Fc-fusion protein in order to
> raise antibodies to the receptor part. So far, so good. I get dozens of
> different antibodies using phage-antibody libraries.
> However, few (or none) of these (and they are not Fc binders !) seem to
> recognise the full-length receptor expressed in mammalian cells, which
> leaves me a bit bewildered. Therefore, the question goes; is it possible
> that the glycosylation machinery in insect cells glycosylates the
> protein in a manner which is dramatically (or at least somewhat)
> different from that observed in mammalian cells ?
> If this is indeed the fact, the differing reactivity of the antibodies
> can be explained by altered epitopes resulting from different
> glycosylations.
>
> Any input appreciated,
>
> Cheers,
>
> Svend Kjaer, Ph.D.
> Dept. of Neuroscience,
> Karolinska Institute,
> Sweden
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