Hi All,
I have done a Protein G purification to purify a recombinant antibody which has IgG1 HC. In the elution phase I was expecting to see a peak that indicates that the antibodies elute. But I could not see any peak. Elution buffer was 0.7% Acetic Acid.
Could you help me to figure out the reason? And is there any possibility to detect and elute my antibody still?
Thanks for your support!
Canan
The first thing to do is to find out if the antibody bound to the protein G resin by looking for it in the flow-through. It may be that the resin is no good and you should get a new resin.
Dear Canan
why are you using acetic acid for elution? i'm not a huge expert on protein but in the most cases i are the 0.1M glicine ph 2.7 or Citric acid ph=3 are used but i never read about acetic acid. did you measure the ph of your elution buffer?
ciao
Manuele
hi cana
you should run the Elisa on hybridoma sup r ascites or culture medium before protein G column purification .If the desired antibody titer was high , you can do:
1-check the the column right activity before use
2- test the new"0.1 M glycine-HCl, pH 2.7" elution buffer
good luck
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