Concentration of antibody - (Jan/14/2007 )

Hi Users.

I have approximately 100mg of monoclonal antibody in approx 80mls PBS. I was hoping to concentrate this down to a much smaller volume but do not want to lose ANY antibody as it is very precious! Any ideas on how best to do this? I was thinking of re-introducing small volumes to the protein G column at a time and re-collecting the eluate. Any better ideas? Thanks in advance!

I think you can concentrate it on the basis of size using concentrating columns (they must work on the basis of size I guess) and centrifugation. May be a bit tedious if you have 80ml as I'm not sure what the maximum volume they hold is. I can find out more if you want.

Ceri

I use a stirred ultrafiltration cell from Millipore. They are a little expensive, and you will lose some antibody. I think that is the easiest and fastest way though.

Thanks guys,

I eventually went with the stirred ultrafiltration option. I'd considered this earlier but thought it might lose too much antibody. In the end I did lose 6mg but from 102mg I didn't think that was TOO bad. Thanks again!

Thanks guys,

I eventually went with the stirred ultrafiltration option. I'd considered this earlier but thought it might lose too much antibody. In the end I did lose 6mg but from 102mg I didn't think that was TOO bad. Thanks again!

wht abt trying spin columns... but if you wnt an easier way.. i would suggest you get a dialysis tube with a 3000 da molecular weight cutoff.. then cover it with PEG. the PEG will absord the water leaving the abs in the dialysis tube.. its fast and easy.. pls be careful not to leave it too long.. as it will absorb all moisture.. good luck..