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Please help me to analyze the experimental result !!!
- Wang
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7 years 2 weeks ago #1
by Wang
Please help me to analyze the experimental result !!! was created by Wang
dear arnoud,
I recently made a kinetic experiment on antibodies, and I'm not sure if my results are reliable. Please help me analyze them.
I use CM5 chip, immobilized anti human IgG Fc, ligand is the antibody of human source, the analyte is protein.
The antibody's dissociatio is very slow, and the HPLC-SEC graph shows that the purity of ligand is 91%, 9% of the polymers, and this will slow down the dissociation of the analyte?
What kind of optimization is needed for this experiment?
Thank you a lot.
I recently made a kinetic experiment on antibodies, and I'm not sure if my results are reliable. Please help me analyze them.
I use CM5 chip, immobilized anti human IgG Fc, ligand is the antibody of human source, the analyte is protein.
The antibody's dissociatio is very slow, and the HPLC-SEC graph shows that the purity of ligand is 91%, 9% of the polymers, and this will slow down the dissociation of the analyte?
What kind of optimization is needed for this experiment?
Thank you a lot.
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- Arnoud
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7 years 2 weeks ago #2
by Arnoud
Replied by Arnoud on topic Please help me to analyze the experimental result !!!
Hi,
The picture is not that good but you have a long dissociation and that is good for a strong interaction. The association time may be a bit longer so the association has some curvature for a better ka estimation.
What you can do is centrifuge the analyte at 15000g for 5 - 10 minutes to remove aggregates / polymers.
I don't think that de polymers are the source of a slow dissociation because 91% is monomer. It seems that you have a strong interaction.
Kind regards
Arnoud
The picture is not that good but you have a long dissociation and that is good for a strong interaction. The association time may be a bit longer so the association has some curvature for a better ka estimation.
What you can do is centrifuge the analyte at 15000g for 5 - 10 minutes to remove aggregates / polymers.
I don't think that de polymers are the source of a slow dissociation because 91% is monomer. It seems that you have a strong interaction.
Kind regards
Arnoud
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