Mesoporous materials have drawn more and more attention in the field of biosensors due to their high surface areas, large pore volumes, tunable pore sizes, as well as abundant frameworks. In this review, the progress on mesoporous materials–based biosensors from enzymatic to nonenzymatic are highlighted. First, recent advances on the application of mesoporous materials as supports to stabilize enzymes in enzymatic biosensing technology are summarized. Special emphasis is placed on the effect of pore size, pore structure, and surface functional groups of the support on the immobilization efficiency of enzymes and the biosensing performance. Then, the development of a nonenzymatic strategy that uses the intrinsic property of mesoporous materials (carbon, silica, metals, and composites) to mimic the behavior of enzymes for electrochemical sensing of some biomolecules is discussed. Finally, the challenges and perspective on the future development of biosensors based on mesoporous materials are proposed.