Affinity assay - Forum


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

Affinity assay 28 Jul 2020 09:30 #1

Hello,

I have a question regarding affinity assay. I am a new user of SPR and have very less or no knowledge about this technique. I am suppose to do affinity assay using Biacore T200 on SA chip. My ligand (biotinylated) is 127KD and analyte is 17 KD. Can you please explain how to setup this assay and then analyse the data?

Cheers!

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Affinity assay 28 Jul 2020 19:23 #2

Hi and welcome,

It seems a bit strange to me that you have to perform experiments without any backup.
The SA chip comes with instructions how to use and you can look at www.sprpages.nl/sensor-chips-intro/biacore-sensor-chips/sa . General information about immobilisation you can find at www.sprpages.nl/immobilization .
For a theoretical Rmax of 250 RU as a starting point you have to immobilize around 2000 RU of ligand (streptavidin-protein).
Once you have some positive binding read the data fitting parts of this website.

I am biased but you could think of buying the SPRpages book.

Kind regards
Arnoud

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Affinity assay 19 Aug 2020 08:53 #3

Hi Gunjita,
I recently got the SPRpages book and I can infact totally recommend to get it, especially for new users (like me) but probably also for experienced users for extra insights.
Best regards, Maria

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