Help with affinity chromatography
jimritt at aol.com
Wed Jan 24 23:21:37 EST 1996
You might try binding your monoclonal directly to the chromatography gel.
I would suggest that you protein A purify the monoclonal antibody first
and then covalently attach it to a commercial affinity gel such as
Pharmacia's CNBR activated sepharose. Some options for eluting your
protein are low pH (glycine/HCl buffer pH2.5), methanol, hi salt (>1M
NaCl) it depends on what your protein is likely to be stable in. After
elution you should get ypur protein back into a friendly buffer.
Pharmacia used to have a useful affinity chromatography guide that was
Good luck - Jim Rittenburg
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