Hi all,
I am trying to express a single chain T cell receptor (i.e. my insert
consists of alpha-chain/linker/beta-chain; together about 700 bases) in
sf9 insect cells using the baculovirus expression system obtained from
Pharmingen. My vector (pAcSecG2T) is designed to enable the protein to be
secreted into the medium. I found the protein (via western blot) in the
cell lysate (the molecular weight was correct) but not in the cell
culture supernatant.It seems as if the protein plus leader got stuck in
the ER due to wrong folding. Has anyone of you who works with the baculo
expression system and tries to express heterodimers like antibodies,TCR`s,
MHC`s,...(as single-chains or alpha and beta-chain separately) ever
encountered a problem like this and what did you do to overcome it? I am
eagerly awaiting some helpful hints to solve my problem.
thanks,
Christian
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Christian Radauer
Institute of General and Experimental Pathology
University of Vienna
AKH 3Q Phone: +43 (1) 40400 5116
Waehringer Guertel 18-20 FAX: +43 (1) 40400 5130
A-1090 Wien
Austria
e-mail: rc150295 at emb1.bcc.univie.ac.at
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