Separation and purification method of recombinant human hyaluronidase

The invention provides a separation and purification method of recombinant human hyaluronidase. The separation and purification method comprises the following steps: (1) enabling a cell culture supernatant containing recombinant human hyaluronidase to pass through a Capto Q anion exchange chromatographic column, and collecting a target elution peak; (2) enabling the target elution peak collected in the step (1) to pass through a Capto pheyI Impres hydrophobic chromatographic column, and collecting a target flow-through peak; (3) enabling the target flow-through peak to pass through a CHT I chromatographic column, and collecting a target elution peak; and (4) enabling the target elution peak collected in the step (3) to pass through a Maxtar SP HR cation exchange chromatographic column, and collecting the target elution peak. Chromatographic fillers used in the separation and purification method are domestic fillers, so that the cost is effectively reduced, and the hyaluronidase with high activity and relative