Problem with SPR data

Hi,
I've recently done SPR to check the binding of a peptide to its target protein (using Reichert 4SPR machine and PEG chip). The graph that I obtaind was not showing a dissociation phase (the curve for the association phase goes up but the dissociation is very slow). When presenting the data, I was asked to fit the RU vs concentration of the peptide then fit the data using a suitable software then demonstrate whether the binding is saturable or not??...…. To be honest I am having a problem understanding what exactly I am supposed to do as I 0 knowledge about SPR and this my first time dealing with SPR data interpretation... I feel sorry asking this but I don't have anyone to help and I was wondering if you have any idea what kind of software I have to use in order to fit the data to single-site binding mode? and how can I demonstrate that the binding is saturable or not...
Once again I am sorry if my question is very naïve I am learning about SPR from scratch..
Thank you so much in advance

Hi HanSue,

Welcome to the SPR community.

To learn the basics, please read first www.sprpages.nl/kinetics and then www.sprpages.nl/sensorgram-tutorial . If your group is new to SPR a good book like the one of this website gives you a very good overview about SPR, methodology and data analysis.

I trust that the SPR instrument has its own control and analysis software. When you have several curves form different analyte concentration injections you can analyse this which gives you the RU vs concentration fit. However, if you have a very low dissociation, you have to be aware that fitting can be difficult. The fit will provide the theoretical Rmax and you can compare that with the obtained maximal response.

I hope I gave you a good starting point.

Kind regards
Arnoud

Hi Han Sue,
If you would like to receive specific guidance on your Reichert SPR system and data analysis, please contact Reichert directly- use the following link www.reichertspr.com/contact/contact-us/

Thank you
Mary