A smart diffraction grating immunosensor based on antigen-responsive hydrogel with enhanced analyte-induced volume changes is developed for highly selective and sensitive detection of human immunoglobulin G (H-IgG). The hydrogel grating contains poly(N-isopropylacrylamide) (PNIPAM) backbones with dual-cross-linking based on the dynamic complexation between pendent goat-anti-human IgG (GAH-IgG) and pendent H-IgG, and the covalent bonding by 4-arm-polyethylene glycol-acrylamide. Upon recognizing free H-IgG in the environment, the pendent GAH-IgG in the hydrogel can form new GAH-IgG/H-IgG complexes with free H-IgG because the binding constant of GAH-IgG to the free H-IgG is much larger than that of GAH-IgG to the pendent H-IgG and thus result in the decomplexation of GAH-IgG/H-IgG complexes with the pendent H-IgG as well as the swelling of hydrogel. The thermo-responsive PNIPAM backbones enable enhancement of H-IgG-responsive volume change of the proposed hydrogel grating via temperature regulation. Moreover, the cross-linker 4-arm-polyethylene glycol-acrylamide provides excellent transparency for the PNIPAM backbones during the volume change, which ensures output of diffracted optical signals with high intensity. With the elaborately designed molecular structures, the hydrogel grating allows highly selective and sensitive detection of [H-IgG] with a detection limit as low as 1.3 x 10(-8) M. This work provides a simple and flexible strategy for developing diffraction grating immunosensors based on stimuli-responsive hydrogels for efficient detection of biomarkers.