Neutralizing Epitope

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dylee's picture
Neutralizing Epitope

I want to know how to find the neutralizing epitope of a virus.
Now we have neutralizing antibodies against the virus in hand and want to do some research on it but we don't know how.
Does anyone had any idea about it?
Thanx very much

protoldo's picture
Hola, I have never done it,

Hola, I have never done it, only in the case of monoclonal Ab.

In the case of polyclonals you know that there are antibodies against some different epitopes. So for monoclonals, I would immobilize antibody in protein A or protein G Sepharose, then I will react it with a virus extract, treated with a specific lysis solution or sonicating , in a column or in a tube.

After washing to eliminate viral proteins and no reactives, I would elute the resin and the eluted peptide would be the epitope. To know the sequence I would send the sample to any proteomic lab. Thinking on it, in the case of polyclonal, probably the proteomic lab could separate the diferent peptides (epitopes) and sequence them, but I'm not sure. 

Good luck