Protein A for surface IgG alignment ?

littgj at wmvx.lvs.dupont.com littgj at wmvx.lvs.dupont.com
Thu Jan 20 19:54:07 EST 1994


In article <CJy934.DvK at crdnns.crd.ge.com>, ebstokes at maxwell.crd.ge.com (Ed Stokes) writes:
>Our group has been coating microtiter well plates with rabbit 
>IgG in an immunoassay. Originally, we were purifying the
>antiserum with protein A affinity chromatography, then
>incubating the wells with purified IgG. More recently, we
>have found that incubating the wells with protein A solution
>first allows us to incubate with antiserum instead of
>purified IgG, eliminating the tedious affinity chromatography.
>
>The plates in this case are polystyrene.
>
>Question: Is anyone aware of a reference which compares
>(a) purified IgG adsorption to polystyrene with (b) protein A
>adsorption followed by antiserum incubation? The literature
>is full of references to protein A coated sepharose beads 
>and gold particles, etc., but I have not been able to find 
>a reference specifically related to microtiter well plate
>coating...
>
>Many thanks
>Ed Stokes
>ebstokes at crd.ge.com
>
I'm not sure about the Protein A/Ab sandwich reference but I've had
considerable experience with Second Ab/Primary Ab sandwichs on 
microplates to do the same thing. The usual (not always) result is
a much higher titer of primary Ab (i.e., you need a lot less), inferring
a more efficient capture. If you want references, I can find you some
(I even patented this for RIA way back in the early 70's)!



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