TCR-Expression using baculovirus

George M. Carter gmc0 at ix.netcom.com
Tue Jul 9 19:54:54 EST 1996


Christian Radauer <rc150295 at emb1.bcc.univie.ac.at> wrote:

>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. 

I ain't no expert. But would these insect cells have the necessary
chaperones needed for correct protein folding?

		George M. Carter





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