The aim of the present work was to develop lysozyme impregnated surface-active nanosponges to maintain its conformational stability and break bacterial cell walls by catalyzing the hydrolysis of 1,4-β-linkages between N -acetyl- d -glucosamine and N -acetylmuramic acid residues present in peptidoglycan layer surrounding the bacterial cell membrane, and for controlling the release of calcium in hypocalcemia condition. Different carbonyl diimidazole cross-linked β-cyclodextrin nanosponges with and without CaCO ₃ and CMC were prepared by polymer condensation method. The surface-active nanosponges were impregnated by lysozyme due to their ability to adsorb protein. Lysozyme impregnated nanosponges had a monomodal particle size distribution of 347.46 ± 3.07 to 550.34 ± 5.23 nm, with a narrow distribution. The zeta potentials were sufficiently increased upon lysozyme impregnation, suggesting stable formulations by preventing aggregation. The in vitro release studies showed controlled release of lysozyme and calcium over a period of 24 h. FTIR studies confirmed the impregnation of lysozyme on nanosponges and encapsulation of calcium in nanosponges. Lysozyme formulation showed promising conformational stability by DSC. It can be concluded that the stable nanosponges formulation is a promising carrier for antibacterial protein and preventing depletion of calcium in antibiotic associated hypocalcemic condition.