Hello everyone, My name is David Vizarraga and I am expressing a single stranded Fv construct. I add a fusion protein like MBP to stabilize the construct and solubilize the protein and this works fine. But when I purify the MBP-ScFv, the whole sample is aggregated (no precipitate). Most likely, S-S interacts between ScFv. How can I avoid that? add some reducing agent like DTT? Or can I add some detergents such as Triton X-100 or Tween20 in buffers? Where to add these agents? and do I need to keep these agents in the buffers all the time? Thank you David
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