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In this chapter, ultrastable synthetic binding pairs between cucurbit[7]uril and its guests are introduced as a supramolecular latching system. The unique features of the synthetic binding pairs including their small size with exceptionally high binding affinity, bio-orthogonality in binding, chemical tractability, scalable synthesis, and robust chemical structures are highlighted with a brief summary of their applications. Other interesting unique features of this binding pair system are also introduced such as delicate control of host–guest binding affinities by treatment with stronger guest molecules on-demand and negligible interferences of the binding properties from endogenous biomolecules, enabling accurate and precise analysis of bioimaging and proteomics.

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