Antibody fragmentation problem

John Russell jrussell at acronet.net
Mon Jun 14 21:12:30 EST 1999

Try adding 6 mg mercaptoethylamine hydrochloride to antibody in 100mM
phosphate pH ~7, incubate 37 deg about 2h.  Desalt into a buffer containing
5mM EDTA, which I understand prevents metal catalysed oxidation back to the
disulfides (I generally feel safer if I first vacuum degas the buffer a few
minutes).  This is from memory of a procedure in a Pierce brochure which came
with the MEA, but should be pretty close.

Nigel Osborn wrote:

> I want to fragment an antibody at the disulfide bond between the two heavy
> chains leaving all the other disulfides intact. This gives a half IgG
> (univalent) with molecular weight around 75kD.
> Does anyone know a method for doing this?
> Thanks.
> Nigel.

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