Thu Apr 12, 2018 2:50 pm by mleipold
Hi Chad,
A while back, I did come up with a way to conjugate MAXPAR polymer to streptavidin or anything else that lacks Cys, or where the thiol-chemistry would screw up the biomolecule (like some of the lectins I published on with Mark NItz). I have attached the protocol: basically, reverse-engineering a thiol-terminated polymer and using Sulfo-SMCC on the biomolecule to attach it. We tested it with some biotin-labeled flow antibodies, and it worked.
The only caveat: I've heard that the biotin-binding pocket of streptavidin monomers contains a Lys which can be derivatized by small molecules (such as Sulfo-SMCC). For a biotin-targeting secondary method like I tested, it's not critical to maintain all 4 biotin-binding sites in a streptavidin tetramer. However, for use in tetramer staining, you'd need to keep all of them for avidity interactions. Evan Newell told me this was one reason why he engineered the mono (or poly) Cys into the mutant streptavidins he made for the CyTOF tetramers.
So, give it a try, but with careful appropriate control experiments. Please report back whether it worked!
Mike