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Big DNA contamination in antibody fraction, HELP ! - (Feb/28/2006 )

Hello everybody

I have a problem : in order to save money rolleyes.gif , 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 wink.gif



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...

good luck