CF2H: a cell-free two-hybrid platform for rapid protein binder screening
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Abstract
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Here we developed CF2H, a rapid and simple workflow for high-affinity binder screening. Our system was designed with the aim of making binder screening simpler, more affordable and accessible to the community. By combining an E. coli lysate-based cell-free system compatible with expression from linear DNA templates to a two-hybrid approach, we constructed a workflow that bypasses tedious cloning, culturing and sequencing steps. Our experimental setup only requires a set of pipettes and a microplate reader for GFP measurements, and its complexity is comparable to setting up PCR reactions.
This is a really interesting method and was a super fun read! I like that not only can it clearly accelerate binder studies, but it can be done in any lab. I'm looking for a use case to try it out already!
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In addition to binding affinity, differences in expression levels, solubility, and stability among binders likely influence the signal output.
Can you interrogate the effect of these different features on the signal output in this method?
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