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How to fit data with mass transport limitations & bulk effects
- bine_-_maya@web.de
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2 years 2 months ago #1
by bine_-_maya@web.de
How to fit data with mass transport limitations & bulk effects was created by bine_-_maya@web.de
Hello,
I'm not sure if my data can be fitted reliably and it would be great to get some feedback. Around 70RU ligand were immobilized and the flow rate was increased to 30µL/min and 50µL/min. But still the reaction does not reach equilibrium and it looks like mass transport limitation and different kinetics or a bulk effect. Samples were diluted in running buffer (at least 1:300). With BSA containing mobile phase a much higher bulk effect was observed. Is it reliable to fit the data with 1:1 binding or is this data not suitable to perform any fitting? E.g. Two state reaction or heterogeneous ligand are theoretically also possible, but I would like to stay with 1:1 even if they look better.
Thank you
I'm not sure if my data can be fitted reliably and it would be great to get some feedback. Around 70RU ligand were immobilized and the flow rate was increased to 30µL/min and 50µL/min. But still the reaction does not reach equilibrium and it looks like mass transport limitation and different kinetics or a bulk effect. Samples were diluted in running buffer (at least 1:300). With BSA containing mobile phase a much higher bulk effect was observed. Is it reliable to fit the data with 1:1 binding or is this data not suitable to perform any fitting? E.g. Two state reaction or heterogeneous ligand are theoretically also possible, but I would like to stay with 1:1 even if they look better.
Thank you
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- Arnoud
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2 years 2 months ago #2
by Arnoud
Replied by Arnoud on topic How to fit data with mass transport limitations & bulk effects
Hi,
Judging the curves the main problem is that the curves are not 1:1 due to heterogeneity (non-specific binding). This is not mass transport. See www.sprpages.nl/data-fitting/troubleshooting/high-buffer-jump
Kind regards
Arnoud
Judging the curves the main problem is that the curves are not 1:1 due to heterogeneity (non-specific binding). This is not mass transport. See www.sprpages.nl/data-fitting/troubleshooting/high-buffer-jump
Kind regards
Arnoud
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