Is there a way to break homodimer scfv and allow it to form a monomer scfv?
Is there a way to convert homodimer scfv into monomer scfv consisting of 2 disulfide bonds at interface of dimer?
You should be able to use exactly the same methods that are applied to antibodies for partial reduction. i.e., incubate with DTT, mercaptoethylamine or TCEP. Since you do not need to preserve some of the architecture of a whole Ab here, you probably have more flexibility, and sodium borohydride could also be used. Normally you would then desalt away the reductant. To prevent re-dimerization, you would need to cap the thiols with an excess of N-ethyl maleimide, for example.
Dear Nick Gee
Will beta mercaptoethanol be able to do this as well?.
If I have a solution of dimers and monomers and I incuvate them with DTT what are the chances that all molecules will form monomers?
Should we dilute it to prevent redimerization?
I need monomers for in vitro testing.
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