Chicken IgY purification

unknown at dl.ac.uk unknown at dl.ac.uk
Mon Jan 10 13:24:23 EST 1994


>Has anybody tried the chloroform/PEG method of antibody purification
>from egg yolk?  Do these antibodies perform well?  The method (Polson,
>1990, J Imm Inv) is easy and gives good yields but the original
>article uses an Ouchterlony assay as the only proof of Ig function --
>I'm a little concerned about possible effects of chloroform on the Abs
>performance in sandwich assays.
>I'd appreciate any feedback from anyone with experience in these
>matters.

Yes, I have used used PEG to purify IgY, although the method I used did
not involve chloroform. The yields were high (approx 5mg protein per yolk,
most of which was IgY). The antibodies work in indirect DAS-ELISA (see
Ed's reply) and in Western blots. I'm currently trying to purify them 
further by HAP liquid chromatography, because of occasional background
problems with some samples caused by excessive lipid (this lipid is harder 
to get rid of than you might think, but thankfully is not present in all
samples. I like the IgY a lot; they work well for me. Another advantage
of the IgY is that they do not cross react with anti-mouse antibodies (as
rabbit Ig sometimes do). 

Ref: van Regenmortel (1982) Serology and Immunochemistry of Plant Viruses
Chpt 4, Academic Press NY.

Please contact me for further information,

Chris

--------------------------------------------------------------------------------
 Chris Rampitsch                       |       RAMPITSCH at BCRSSU.AGR.CA
 Dept. Plant Science, Univ. of BC      |       Phone (604) 494-7711
 Vancouver, British Columbia           |       Fax   (604) 494-0755
 Canada V6T 2A2                      _/\_
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