Ab purification problem
AEVDOKIMOZ at cinci.rr.com
Thu Oct 17 22:40:05 EST 2002
> Another possibility, although not very likely, is that epitopes
> normally hidden in the native protein are much more
> immunogenic than the ones exposed. In which case, particularly
> for late, mature bleeds, majority of Ab in the serum will not
> bind to the column made with native antigen. They will work
> perfectly fine on Western because antigene there is denatured
> and "crucified" on membrane. This kind of thing happens
> all the time with moAb. That kind of thing can account for your
> low yeild, too.
That's why I asked if he knows at least some of the actual epitopes.
Depending on the protein, it may be possible to find surface continuous
epitopes, have these peptides synthetically made, and to raise antibodies to
them specifically, which would make purification considerably more reliable.
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