Horseradish peroxidase (HRP)-catalyzed hydrogelation has attracted much attention owing to the ease of handling, high biocompatibility, and processability. This review summarizes recent developments, including cutting-edge research into the use of HRP-mediated hydrogelation toward biomedical, biopharmaceutical, and biofabrication applications. From the viewpoint of polymer chemistry, the basic chemistry behind hydrogelation, the structure-property relationship, and hybridization of multiple materials by using HRP-catalyzed hydrogelation are summarized. From the chemical engineering perspective, strategies for controlling hydrogelation kinetics, hydrogel characteristics, and hydrogelation processes are summarized and discussed in detail. Strategies for obtaining biomaterials for medical and pharmaceutical use, and biofabrication for tissue engineering and regenerative medicine emerge by unifying the aspects of HRP-mediated hydrogelation.