Big DNA contamination in antibody fraction, HELP ! - (Feb/28/2006 )
I have a problem : in order to save money , I tried to purify by myself my precious antibody from hybridoma supernatant. I used the MabTrap Kit from Pharmacia (G protein; OK, the expiration date has been reached 2 years ago...) I checked the OD at 280 and 260, and surprise, I have nearly the same absorbance (e.g. : A260, 1340; A280, 1449; A320, 1330)
So Isuppose that I have strong DNA contamination ?
I have 2 questions :
1/ how to improve my purification
2/ If impossible, how to free my antibody from any DNA ?
3/ it would be wonderful if you had an unexpensive solution...
thanks a lot
I have never tried to purify my own ab but to determinate it from DNA, I would try to sonicate it or to give a DNase to the solution...