We just pulled a few interesting scFv (single-chain Fv) antibodies out of a phage display library. Now we want to produce a soluble antibody from these clones. While the phagemid allows for direct expression of tagged, soluble scFv in E. coli, we are also interested in reconstituting the scFv fragments in a way that ordinary, species-specific anti-Fc secondary antibodies will recognize them.
I'm aware that expression of full IgG is possible only in a eukaryotic system. I found a few protocols to make "humanized" IgG starting from Fab or scFv, but we want mouse antibodies. We are looking for a convenient vectors to fuse scFv to the murine Fc domain. Anyone know of a source?
Dear Alexandra,
please look at the vectors from Invivogen. It may be possible that these constructs will help you in solving this problem.
Best regards, Juergen
I agree with Juergen, the vectors from Invivogen are excellent to solve your problem.
Best regards,
Gestur
Dear Alexandra,
I agree with Juergen, the vectors from Invivogen are excellent to solve your problem
Thanks everybody, for pointing me in the right direction! At least we know more precisely now, what we need. Invivogen plasmid is perfect, but not sure we can afford it at this point. I will check out all the alternatives, respectively get in touch with the people suggested.
Thanks again! :)
it's fairly easy to reformat them using PCR splice overlap extension.. especially if you have a nice program like Vector NTI to help you design primers
@ Richard: Yes we are aware of that. But time is an issue, in a production unit like ours. At this point we wouldn't want to put all the effort in, making our own Fc fusion vector. It's a pilot project. We're not yet sure whether or not making antibodies is going to become a permanent part of our portfolio, so... Trying to go the quicker way ;)
Thx
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