Biotin-tagged molecule immobilisation

Hi all,

I have read that successful biotin-tagged molecules immobilization need to reach a stable plateau.
Is it possible to reach a stable plateau also if you want a low immobilization level?
Thanks!
spruser

I'am not sure I understand your question.

The avidin-biotin binding is a very strong binding. The binding is strong enough to be considered stable for your purposes.

When binding your biotin-tagged protein to a avidin sensor surface the response after immobilization must be stable even if you have a low immobilization level.

You can block the remaining avidin sites with a biotin injection and stabilize the surface with some high salt pH 8.5 injections (if possible with yor ligand).

Arnoud

Thanks for your reply Arnoud,


Do you have access to BIAapplications Handbook from Biacore? At the end of the Streptavidin chapter (Page 4-26) it says that successful binding to SA surface means that the immobilization curve reach a stable plateau.
spruser