Forum :: Increasing sensorgram in dissociation phase


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

TOPIC: Increasing sensorgram in dissociation phase

Increasing sensorgram in dissociation phase 30 Jan 2016 12:22 #1

I am working on a project determining binding kinetic of an antigen (TNF) to
an antibody (Etanercept). The sensorgram became increasing in the dissociation
phase, I have attached the results. Does anybody have any idea about
the probable reason that may be related to this problem?
It is noteworthy to mention that I have desorbed, sanitized and
supercleaned my instrument before.

20151118_184316.jpg

Increasing sensorgram in dissociation phase 01 Feb 2016 09:02 #2

Hi ramin,

Cleaning the instrument may give some extra drift. This will become extra visible when having long dissociation times (as with the curves you show).
Normally I let the system on standby overnight after the cleaning procedure to minimize drift.

It is also a good idea to have multiple dummy runs before your experiment and have some blanks in between the analyte injection to perform some double referencing. If drift is persistent, double referencing can possibly compensate.

You may want to read the experimental strategy page (www.sprpages.nl/experiments/strategy) for other experimental set-ups. Since you have long dissociation times it can be beneficial to use an other strategy.

Kind regards

Arnoud
Last Edit: 01 Feb 2016 09:03 by Arnoud.

Increasing sensorgram in dissociation phase 04 May 2016 01:33 #3

Question; did you run the higher concentration analyte injections before the lower concentrations? You could have drift due to the fact that the surface has not been completely regenerated.
Thanks!

Increasing sensorgram in dissociation phase 04 May 2016 19:31 #4

The experiment started with lower analyte concentrations. The regeneration condition is ok.
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