I am trying to remove selectively an enzyme from a crude extract using
binding to an antiserum. In short, I hope to bind antibodies to the
enzyme and then remove this by binding to beads coated with protein A
But it doesn't work. My extract still has the same amount of enzyme
activity. I tried a number of approaches:
1/ first add the antiserum to the extract, incubate overnight at 4C,
then add beads with protein A/G
2/ titrate antiserum in the above experiment
3/ first bind serum to beads for 4 hours, then wash and incubate with
extract overnight at 4C.
Has anybody ever tried to do a similar experiment, have seen this
published, or is anybody able to give me advise?
Thanks in advance
Koen De Smet
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Imperial College School of Medicine at St Mary's