I am very new to SPR. I have been using CM5 chip (BIA 2000) and immobilized the ligand. The ligand is a hexamer. Literature suggests that one molecule of analyte binds to the hexamer or 6 molecules of ligand. Is it possible to analyze this using SPR. What precautions should I take before passing the analyte.
Sorry for the late answer and thanks a lot for you prompt reply. Yes, My ligand is a hexamer which binds to single molecule of analyte. (Reference : Alternating translocation of protein substrates from both ends of ClpXP protease www.ncbi.nlm.nih.gov/pmc/articles/PMC126282/)
Could you please guide me how to fit the data with existing model or precautions which are clearly needed with this kind of system.
When the ligand is a hexamer, binding only one analyte molecule, in SPR you model that as a 1:1 interaction. In general, the ligand is treated as if it is monovalent. For instance an IgG is bivalent but from the SPR perspective both binding sites are independent. In your case you need a hexamer to form one binding site and therefore you should model is as a 1:1 interaction.