Protocol Online logo
Top : New Forum Archives (2009-): : Protein Expression and Purification

Weak affinity chromatography - (Apr/20/2012 )

I am looking to enrich serum IgM for specific antigen utilizing weak affinity chromatography (wac). Protocols that I have encountered for WAC utilize HPLC, which I do not have access to. Is it possible to do this by FPLC or gravity with similar resolution?Any advice for enriching specific serum IgM with incredibly weak affinity in a gentle (nondenaturing) approach?


weak can mean "less specific"; you may use hydrophobic interaction chromatography where you can gear binding by salt concentration or pH...

-Inmost sun-

if you can find the same packing phase as the hplc method then you can run fplc or/and conventional lc. fplc resolution should be similar, conventional may exhibit more band spreading due to slower elution.