[Immunology] IgM purification

Aaron Wilson via immuno%40net.bio.net (by aaron.wilson from gmail.com)
Tue Apr 17 18:44:52 EST 2007


Hello Dr. Preston,

  I am a graduate student at Carleton University in Ottawa working
with Dr. J.David Miller and I happened to stumble across a suggestion
you had made regarding IgM purification with gel filtration (many
years ago), and I was wondering if I could seek some advice?
We are attempting to purify IgM antibodies from mouse cell
supernatant,  (not quite at the MAb ascites stage yet) and so far we
have being encountering large antibody losses as well as undetectable
increases in purify of the sample using the affinity chromatography
approach.

You had mentioned that you had used gel filtration had had excellent
recovery and high levels of purification.  We currently use the S300
resin, generally eluting our allergen protein samples with 0.15M NaCl
50mM Tris/HCl, pH7.5 buffer.  I was just wondering if you had any tips
or suggestions for us utlizing this procedure to purify our IgM
antibodies from the cell supernatant?

Any comments or suggestions would be greatly appreciated.

Best Regards,
Aaron Wilson



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