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How do i know when the fit is good?
- Zackarias
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3 years 3 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?
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?
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- Arnoud
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3 years 3 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.
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