I assume that you do not have any SDS in your sample.I have not tried mABs
purification by RP-HPLC,but I have been pretty successful using a
C18-4000A column.The column was from Synchrom.I know that they have
acquired by an
outfit in either Illinois or elsewhere in Ohio,and at least two years ago
they carried the columns,under the same name,
Synchropak.The type of protein which I tried to purify was more
glycosylated than any immunoglobulin.and a bit smaller.
It is possible that 1000A porosity will do,but the usual 300A,definitely
won't cut it.
"Wei Zhang" <WXZhang@genetics.com>@aecom.yu.edu> on 11/07/2000 09:22:30 AM
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Subject: RP-HPLC for mAb
Item Type: Task
Start Date: 11/07/2000 (Tuesday)
Due: 11/07/2000 (Tuesday)
We are developing RP-HPLC methods for mAbs. We've tried C18, C4, -CN and
phenyl columns from different vendors, none of them gave good results in
terms of peak shape and resolution. We saw some minor bands in SDS-PAGE and
are trying to use RP-HPLC to resolve them. Could you please share with us
your experience with RP-HPLC of mAbs or other large proteins? Thanks a lot!
Wei Zhang, Ph.D.
Genetics Institute
wxzhang@genetics.com
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