How topurify and measure stability of mAb IgM - (Dec/02/2009 )
Hi Fatemeh,
Have you had any luck purifying your IgM MAb? I too am trying to purify an IgM and it is proving to be quite challenging! I am using the GE -HiTrap IgM purification column and am trying to purify from supernatant harvested from the CELLine CL-350 flask. I have concentrated ny harvest before loading on the column but seem to get a very low MAb yield. Any advice would be much appreciated.
-MouseMab-
MouseMab on Apr 14 2010, 10:18 PM said:
Hi Fatemeh,
Have you had any luck purifying your IgM MAb? I too am trying to purify an IgM and it is proving to be quite challenging! I am using the GE -HiTrap IgM purification column and am trying to purify from supernatant harvested from the CELLine CL-350 flask. I have concentrated ny harvest before loading on the column but seem to get a very low MAb yield. Any advice would be much appreciated.
Have you had any luck purifying your IgM MAb? I too am trying to purify an IgM and it is proving to be quite challenging! I am using the GE -HiTrap IgM purification column and am trying to purify from supernatant harvested from the CELLine CL-350 flask. I have concentrated ny harvest before loading on the column but seem to get a very low MAb yield. Any advice would be much appreciated.
Hi MouseMab
Sorry for very late reply, I hope you could get very good results by now. I should say I couldn't yet. I use the same column as yours but still no desired results. May I know what protocol you follow? My protocol is explained in my previous posts.
Regarding concentration, have u checked your IgM concentration after concentrating? If you are using concentrating filters, sometimes it adsorbed the protein you should be careful in choosing the concentrator avoiding the low yeild of concentrated antibody. Ammonium precipitation may be better choice if it doesn't affect IgM activity.
Good Luck,
fatemeh
-fatemeh-