Purifying Chicken IgY

RYBICKI, ED ED at micro.uct.ac.za
Mon Jan 10 08:32:11 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.

Re: Rybicki, 1979 (MSc Thesis, Univ Cape Town), and Rybicki and von 
Wechmar, 1981; Virology 109: 391-402).  You can use them in ELISA, 
but it is difficult to conjugate them with Alk Phos (don't know 
why), so used a compound sandwich technique or indirect (eg: 
rAb-virus-cAb-rabbitAbantichickenIgYAlkPhos or virus-cAb-rAbAP).  
Worked very well indeed.  Think we once used them for westerns too 
(had lots of alkphos-conjugated rabbit-anti-chicken IgY around, 
seeing as Polson was working here at the time) and they worked fine. 

Hope this helps,

  ____________________________________________________________________
 | Ed Rybicki, PhD             |                                      |
 | (ed at micro.uct.ac.za)        |      "Lord, won't you buy me         |
 | Dept Microbiology           |          A Mer-ce-des Benz           |
 | University of Cape Town     |     My friends all have Porsches     |
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