By Anonymous on Monday, February 12, 2001 - 09:28 am:
Hello,
I am trying to detect various degradation products of monoclonal antibodies using HIC.
I am using a TSK-Butyl-NPR column, with a
gradient of 2M ammonium sulfate from 100 to 0%
over a 30 minute duration. The column temperature
is 40C. The peaks observed are unfortunately
not well resolved. Could someone please
suggest what I could do to improve the resolution
(any other columns/ salt etc)?
Thanks!
By tom jupille on Wednesday, February 28, 2001 - 05:24 pm:
Depejnds on how poorly resolved or separated your peaks are. If the peak shape is OK (sharp, symmetrical peaks) but the selectivity (peak spacing) is borderline, you might try a shallower gradient (longer time or narrower salt range).
If the peaks are reasonably separated but excessively broad or poorly shaped, you might try changing the temperature by 5 degrees either way (on the possibility that you've got an equilibrium mixture of different conformers under your present conditions).
-- Tom Jupille / LC Resources
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