problem with elution from hydrophobic column

blatch at uctvax.uct.ac.za blatch at uctvax.uct.ac.za
Tue Mar 9 07:03:31 EST 1993


PROBLEM WITH ELUTION CONDITIONS FOR A HYDROPHOBIC COLUMN:

We are trying to purify a protein using a hydrophobic material called FRACTOGEL
TSK BUTYL-650 (M) produced by Merck. The matrix consists of butyl residues
linked by an ether link to a polyvinyl support matrix. Our protein sample will
stick to the matrix at approx 0.8 M ammonium sulphate but under no
circumstances will any proteins elute from the matrix even if we use distilled
water. Note our protein sample will not stick to the matrix in the absence of
salt. So we are dealing with what seems to be an irreversible binding process.
What are we doing wrong?  Must we just change to a completely different
hydrophobic moiety? Any suggestions will be greatly appreciated. Please send
suggestions to me or Neil at micro.uct.ac.za



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