Forum :: Multivalent Binding


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This forum is intended for questions about kinetics, Surface Plasmon Resonance and the instruments related to these techniques.

TOPIC: Multivalent Binding

Multivalent Binding 05 Feb 2016 04:28 #1

Hello All,

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.

Thanks a lot..
Last Edit: 05 Feb 2016 04:28 by Deeptil.

Multivalent Binding 05 Feb 2016 10:08 #2

Hi,

I am a little confused. Do you mean that one ligand molecule (a hexamer) can bind 6 analyte molecules or that to need the ligand hexamer conformation to bind one analyte molecule.

Can you quote the publication?

Kind regards
Arnoud

Multivalent Binding 24 Feb 2016 09:50 #3

Hello,
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.

Thanks a lot

With Regards,
Deepti
Last Edit: 24 Feb 2016 09:50 by Deeptil.

Multivalent Binding 24 Feb 2016 10:30 #4

Hi Deepti,

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.

For some extra information on the parameters look at
www.sprpages.nl/data-fitting/models

Kind regards
Arnoud
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