What conditions do you use when cleaving DDE-based linkers in protein bio-conjugates (time, temperature, hydrazine concentration?) Are there linkers which can be cleaved in milder conditions and/or cleave more efficiently than DDE/hydrazine? We can't use S-S linkers b.c. the protein does not survive purification in the absence of reducing agents. Reviews on cleavable linkers (e.g. a recent one at 10.1039/C8CS00676H) seem to define "mild" as 25-37C for hours which would not work for us. The only alternative I can think of is to use a TEV-cleavable peptide (4C overnight)
Upon further reading, it appears that nitrophenyl-based photocleavable linkers might offer more efficient/milder release conditions since I could do it at 4C (or even at -20 if I use 50% glycerol). But I wonder if there are any caveats in using PC linkers since I was not able to find many papers that use PC linkers. The following paper claimed 73% elution:
10.1002/mabi.201800104
- Badges
- Science topic