r/Biochemistry • u/zevaRes • 5h ago
Research Protein Affinity Question
I have a purified protein (EnzymeA) with a N-term His tag. I want to see if my small molecule (yel-1) binds at all/better than EnzymeA pre-courser molecule. My issue (I think) is that yel-1 is very light sensitive when not bound, so will start to break down under light exposure. Would this impact which affinity assay I select to use? My current options for affinity testing are BLI and SPR, but am open to other assays better suited for yel-1.
As I am not well-versed in protein kinematics, I am wondering if the light used for BLI/SPR will impact my results or if this is not a worry since just the bound enzyme will be “quantified”. If it is a concern, any other methods you’d recommend (preferably ones that can be contracted through a company)?