I am looking for some protocol to purify a MAb IgM isotype from ascites fluid. I have found purifying methods to isolate IgM from plasma or serum and MAbs IgM but from culture supernatant. Please, if somebody knows about it, I need this information
From the experience in our lab we usually use this simple protocol for the purification of IgM monoclonal antibody from serum-containing cell culture medium:
1) Precipitate the IgM antibody with 50% Ammonium Sulphate.
2) Dissolve the pellet in 0.1 M Ammonium Bicarbonate and load it on the column with Sephacryl 200. The first peak eluted from the column will be highly purified IgM antibody (not 100% purity). There are not many proteins in blood serum (or ascites) which have MW about 1,000 kDa as IgM (pentamer).
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