The D-glucose imprinted core-shell nanosphere with an average size of approximate to 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 biomimetc model system with a well-defined morphology, high surface area, and variable biding affinity through a change in temperature.