Hi Jeff

I think you need to be clear about the differences between binding and neutralisation with regard to your snake venom immunogen. The total chicken antibodies that you have will contain a large percentage of non-specific antibodies (those which the chicken normally produces to combat infections etc to keep it alive and healthy). Additionally it will contain specific binding antibodies against your venom immunogen. Of these specific antibodies many will bind to non lethal components within the venom (snake venoms contain a complex mixture of digestive and toxin proteins) or to non-neutralising epitopes of toxin molecules or with such a low binding avidity (affinity) so as to be non-neutralising. A relatively small fraction will bind with a high enough avidity (affinity) to critical regions of toxin molecules to either directly block the active site or cause subtle structural changes resulting in the loss of activity and therefore neutralisation.

To complicate things further many snake venoms contain a cocktail of toxins so purifying the single main toxin and immunising will produce a potent single toxin neutralising antisera which may fail to neutralise the lethal effects of the whole venom (a good example of this can be seen in this reference plus methods to measure specific binding antibody levels: https://www.researchgate.net/publication/15474521_Experimental_evaluation_of_ovine_antisera_to_Thai_cobra_%28Naja_kaouthia%29_venom_and_its_alpha-neurotoxin?ev=prf_pub

In vitro neutralisation assays such as the following may also be an option: https://www.researchgate.net/publication/13109097_The_effects_of_specific_antibody_fragments_on_the_%27irreversible%27_neurotoxicity_induced_by_Brown_snake_%28Pseudonaja%29_venom?ev=prf_pub

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I am totally agree with Dr Russell G A Jones

To quantify the specific antibodies against the venom...you must develop a specific ELISA for that.

If you want that

i can help you and send you all steps for that

That would be great Naoual, we have been developing a specific ELISA trying to determine how much specific antibodies will effectively neutralize the venom.

The binding of an antibody doesn´t mean neutralization. To assess a "neutralization" in vitro test, you should know the primary and conformational structures in your toxin(s) which should be blocked to avoid its toxic action. Only in this circumstamces you could suppose that your in vitro results are congruent with neutralization assays. In example the binding of antibodies to a PLA2 or inclusive to block its enzymatic activity, not alwas mean that this antibody can neutrailze the toxic activity. The same for other toxic components of venomous animals.

Dear Jeff, 

Here is my email: [email protected]

The difficulty is in that there are several toxins present.  I have also read that it is possible for more than one antibody to bind to a single antigen.  What I am trying to do is quantify the amount of specific antibodies present in a sample of purified avian antibodies. 

You could use small scale affinity chromatography to quantify the amount of venom or toxin specific antibody.

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We have thought about that but the issue becomes which antigens from the venom are binding to the column?