The IFNg secretion assay uses 2 antibodies (one surface marker and one interferong specific) that are conjugated together. Is it possible to take 2 urified antibodies and do the same conjugation (preferably without having to make recombinant antibodies..)?
This paper (link attached) seems to have done this with IL-21 (see supplemental data) however, I am having a hard time finding how they conjugated Streptavidin to the surface Ab as this would mean the streptavidin or some component was fluorescently conjugated to enable downstream sorting of live cells. At the moment, i cannot find any streptavidin conjugation kit that contains a fluorophore. If you can tease it out, please let me know.
http://www.sciencedirect.com/science/article/pii/S1074761315005373
Best Wishes,
Ane
Indeed, this is what we did in the article mentioned above. We used anti-MHC Ab conjugated to streptavidin, and then used a biotin-conjugated anti-IL21 Ab to make an anti-IL21 capture Ab tethered to the cell surface by the biotin-streptavidin bond. For the downstream flow application you mentioned, we used a different anti-IL21 Ab, conjugated to PE, in essence making an IL21 sandwich. Hope this helps.
Sorry didnt get the notices for these answers, thankyou both very much! Good luck Ane!
You could also have a look at the GlyCLICK conjugation from Genovis for site-specific labeling at the Fc glycans. https://www.genovis.com/products/glyclick/
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