I want to dissociate virus bound to antibody onto the Protein G sepharose high performance columns. Can anyone suggest a simple protocol which will enable me to separate them while keeping them intact? I came across some suggestions, like using KSCN or 0.1-0.4M MgCl2. I don't want to use any kit for the purpose. Any help will be appreciated. Thanks in advance.
Please find the attached protocol using lower pH as the basis for release by
Christina Milburn 2/13/97
I have used similar protocols
Hi! Just guessing now from my side, as I have no experience with that particular problem and I don't know with which virus you are working and how well it supports low pH or detergents and how well your antibody binds the virus - is it feasible to dissociate the virus via dilution of the complex (i.e. via its association constant)? Please keep me informed concerning the solution of your problem!
If you want to dissociate just virus from the complex, this process depends affinity of antibody. If your antibodies have low affinity, a little bit pH lowering is sufficient for dissociation. But, high affinity is a big problem. Because it needs big pH changes. Big changes of pH can affect protein G-antibody bounds. This is a big problem. Please let me know more information about your problem.
Hi! Concerning pH change, you have to be careful not to induce virus uncoating (if you want to obtain intact virions)...
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