Change in elution time with protein concentration
Posted: Fri Feb 26, 2010 11:15 am
Good morning,
I saw something today which I have not seen before and haven't heard of. Perhaps someone with more experience can advise?
I am injecting an intact protein on a C3 column, 300A pore (for analysis only, hence not HIC or ion exchange), in a gradient of acetonitrile in 0.1% TFA. Injecting the protein from its storage buffer (low buffer strength, 100mM sodium chloride) gives a reproducible, symmetrical peak. However, diluting the protein by 10x in the same buffer before injection of the same volume gives a peak of a tenth of the area, but the elution time is quicker.
Is this normal?
With thanks,
D.
I saw something today which I have not seen before and haven't heard of. Perhaps someone with more experience can advise?
I am injecting an intact protein on a C3 column, 300A pore (for analysis only, hence not HIC or ion exchange), in a gradient of acetonitrile in 0.1% TFA. Injecting the protein from its storage buffer (low buffer strength, 100mM sodium chloride) gives a reproducible, symmetrical peak. However, diluting the protein by 10x in the same buffer before injection of the same volume gives a peak of a tenth of the area, but the elution time is quicker.
Is this normal?
With thanks,
D.