Ion Exchange Chro/HPLC with 6M Guanidine or Some Detergent?

newera at newera at
Sat Mar 8 08:59:17 EST 1997

Do not 6M ganidine HCl, 8M urea or some zwitter-ionic detergent make any difference to cation exchange chromatography or affinity chromatography?

My protein tends to be aggregated during some purification steps such as dialysis or concentration with a Centriprep. In the midst of misfortune, my protein has no cysteines and seems to be easily renatured; so I plan to add 6M guanidine HCl/8M urea and 0.5~2% or more zwitter-ionic detergents such as CHAPS, sarkosyl to my current buffer system(20mM Na Phosphate, pH 6.8). 
I am deeply concerned that they severely interfere with my protein's binding ability to BioRex70 cation exchange resin, BlueSepharose affinity chromatography resin, to which my protein seems to quite tightly bind since it begins to elute at around 0.5M NaCl, and C18 reverse phase HPLC.

Thank you very much.

Key word:
6M Guanidine HCl/8M Urea
Cation Exchange Chromatography
Affinity Chromatography
Reverse Phase HPLC

email 	: newera at
address :
  Lee, Ji Hyun
  Laboratory of Physical Pharmacy(Prof. Lee, Bong Jin)

  Seoul National University
  College of Pharmacy		  
  Shinlim-Dong, Kwanak-Gu
  Seoul 151-742, Korea.

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