Immuno-affinity Chromatography

Basavaraju Shankarappa bsh at MED.PITT.EDU
Fri Dec 11 15:40:53 EST 1992

> I have purified IgM antibody from my culture media by Sephacryl 300.  I have
> also tried to couple this antibody to CNBr-activated Sepharose 4B.  Antibody
> binds to the beads, but it loses its activiy and leaks with the eluting buffer
> (0.1M Glycine, pH 2.5).  How do I get around these problems?
> Hung Nguyen
> Biology Dept., Boston College

	I had used affi-gel matrices for immunoaffinity purification of
antigens a while back.  About two years back BioRad started marketing
affigel HZ which is supposed to bind to the antibody at its Fc region
through carbohydrate moeities (?).  Is it worthwhile to try it?
Has anybody tried it.  The company's graphs show a considerably increased
yield.  May be it is better to trust results from some lab.  
I do not see any reason why IgM should not bind in the same way.
Raj Shankarappa
bsh at
Univ of Pittsburgh

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