The invention discloses a method for highly purifying kallikrein. The technical scheme adopted is that the method comprises the following steps: using hog pancreas as raw materials for extraction and depositing to prepare acetone powder; flexibly using combined traditional and modern protein purification technologies such as acetate extraction, acetone deposition, acetic ether degreasing and dehydration, sodium chloride-ammonia water and acetone collaborative impurity removing, ion exchange resin DEAE-agarose gel substep elution, and highly purifying the kallikrein. The method is short in production period, has good chemical and thermal stability, is low in production cost, and is very suitable for large-scale production.