Optimization of Condition for Immobilization of Trypsin and Application of Immobilized Trypsin in Purification of Aprotinin

Objective To optimize the condition for immobilization of trypsin and apply the immobilized trypsin to the purification of aprotinin. Methods Immobilize trypsin using SBA-15 as carrier and glutaraldehyde as crosslinking agent and optimize the condition for immobilization. Isolate and purify aprotinin using the trypsin immobilized under the optimized condition as affinity adsorbent. Results The optimal glutraldehyde concentration, sample loading quantity, temperature and time for immobilization of trypsin were 0. 8%, 3. 0 mg, 35℃ and 3 h respectively. The activity of trypsin immobilized under the optimized condition reached 15. 6 U / mg. The purification fold of aprotinin with the immobilized trypsin was more than 420, and the recovery of activity was more than 80%. Conclusion The condition for immobilization of trypsin was optimized, and the purification effect of aprotinin with the immobilized trypsin was satisfactory.