glycosylation in insect cells ?

Svend Kjaer svend at CAJAL.MBB.KI.SE
Tue Jun 29 12:44:12 EST 1999


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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