Has anyone had experience with purification of biotinylated proteins from free
biotin by dialysis?
I have been using EZ-Link-Sulfo-NHS-LC-biotin( from Pierce) to biotinylated mAB
and other proteins. After biotinylation, I use excess amount of Lys to stop the
biotinylation reaction. I tried to use 6,000-8,000 MWCO molecular membrane to
dialysis the biotinylated protein against 1x PBS buffer. But no success. By
LC/UV/MS, I can still see a lot of free biotin in the dialysised sample.
I can only get rid of the free biotin by Protein A purification or SEC method.
Is this due to the strong non covalent binding between the free biotin and the
protein?
Thank you for your help.
Rong-Rong Zhu
BASF Bioresearch Corp.
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