Help with affinity chromatography

Jim Ritt 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
free.

Good luck  -  Jim Rittenburg



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