Synthetic modification of commercially available cyclodextrins offers a straightforward way of increasing the diversity of host-guest interactions. However, modifications of cyclodextrins have focused largely on persubstitution with a small set of substituents; random substitution, which yields products not suited for rigorous comparison with calculations; and mono-substitution of only the more synthetically accessible 6' hydroxyls of the smaller primary face (Figure ). Mono-substitution of the less reactive 2' and 3' hydroxyls of the larger secondary face has been regarded as relatively challenging. We recently addressed this with synthetic approaches to modify not only the 6' but also the 3' position with a variety of substituents, and to simplify purification of the desired products. We hope ultimately to generate a dense matrix of thermodynamically characterized host-guest interactions (Figure ) that will be of ongoing value to researchers seeking to test or parameterize computational models of binding and thus to advance the accuracy of computer-aided drug design. As a next step in this direction, we report here the binding thermodynamics, measured with isothermal titration calorimetry, of two guest compounds (Figure ) with β-CD and eight diverse monosubstituted β-CD derivatives . The β-CD substituents include a steroid nucleus; cationic, neutral and anionic groups; an aromatic group; and a cysteine-containing peptidic chain. The conformational preferences of selected complexes were further characterized by 2D NOESY NMR experiments. These cases were adopted for use in the SAMPL7 blinded prediction challenge detailed in other papers of this special issue.