SPR-Pages

pagelogo
back
How do i know when the fit is good? - SP
back
This forum is intended for questions about kinetics, Surface Plasmon Resonance and the instruments related to these techniques.

How do i know when the fit is good?

  • Zackarias
  • Topic Author
  • Visitor
  • Visitor
3 years 2 months ago #1 by Zackarias
How do i know when the fit is good? was created by Zackarias
Hi, I am a PhD student who is new to SPR without anyone to really ask questions to so i was hoping i could perhaps get an answer here.

I have these two ligands that i am comparing and running analyte over and i want to compare the binding.
As the fit is better in the lower concentrations does that mean that i am mass transfer limited in the higher concentrations?
Is the fit still good in the association as i do have curvature?
During dissociation the baselines does not return to zero, how much will this affect the Koff and is this expected to have some binding still left?
Attachments:

Please Log in or Create an account to join the conversation.

  • Arnoud
  • Visitor
  • Visitor
3 years 2 months ago #2 by Arnoud
Replied by Arnoud on topic How do i know when the fit is good?
From the inset I deduce that the sensorgram is double referenced. Despite this, there is clearly some non-specific binding which proportional to the analyte concentration used. Also in the association there is a sign of non-specific binding where the measured curve crosses the fitted curve. The dissociation fit is not following the curves and therefore the calculated values are not very reliable. I don't think you have to worry about mass transfer here. On the other hand it seems that the analyte concentration is still below KD. Please study www.sprpages.nl/sensorgram-tutorial/a-curve and look at the curve spacing.

Please Log in or Create an account to join the conversation.

Moderators: ArnoudArnoud
Powered by Kunena Forum