重组羧肽酶B的稳定性研究

目的筛选重组羧肽酶B(rCPB)酶液的保存方法以及rCPB的冻干保护剂。方法将添加和不添加甘油的两组rCPB酶液置于不同温度下保存,定期测定活力;在rCPB酶液中加入不同浓度的不同保护剂,比较冷冻干燥后的活性保留率,并从赋型性和溶解性方面评价冻干保护剂的作用。结果 4℃下,含25%甘油的rCPB酶液在6个月内活力保持不变;添加3%甘露醇和5%海藻糖的rCPB冻干品的酶活保留率分别为102%和100%。结论甘油能显著提高rCPB酶液的稳定性;从冻干保护效果和冻干品外观来评价,3%甘露醇可作为rCPB冻干过程中的最佳保护剂。

Study on stability of recombinant carboxypeptidase B

Purpose To investigate the storage condition for recombinant carboxypeptidase B solution and to screen the reagents to protect rCPB in the process of lyophillization.Methods RCPB solution containing 25% glycerol or without glycerol were stored at-20 ℃,4 ℃ or room temperature,respectively,to be investigated for the activity at different durations.Trehalose,mannitol and sorbitol were selected to protect the activity of rCPB from losing during lyophillization,at the concerntration of 1%,3% and 5%,respectively.Results The activity of rCPB with addition of 25% glycerol kept unchanged at 4 ℃ within six months.No activity of rCPB was lost when 3% mannitol or 5% trehalose was used as additives for rCPB lyophillization.Conclusion Glycerol was helpful for the storage of rCPB solution and no activity was lost within 6 months when kept at 4 ℃.3% mannitol was selected as the best protector for the lyophillization of rCPB and there was no activity losing in the process of lyophillization and a good appearance was got.