Summary: The D ‐glucose imprinted core‐shell nanosphere with an average size of ≈60 to 80 nm showed a significant preference for the binding of D ‐glucose than the non‐imprinted core‐shell nanosphere. Depending on temperature, the binding site in the shell with N‐isopropylacrylamide oligomer underwent a significant change in binding affinity. In addition, the D ‐glucose imprinted core‐shell nanosphere showed a two times higher affinity for D ‐glucose than L ‐glucose, suggesting chiral recognition of the binding site. The core‐shell nanosphere reported here is a good biomimetic model system with a well‐defined morphology, high surface area, and variable binding affinity through a change in temperature.