摘要
目的制备可溶性重组人成骨蛋白-1(rhOP-1),并尝试制备注射制剂。方法rhOP-1工程菌经发酵表达,收集菌体,提取包涵体,用SP-FF阳离子层析柱纯化。取纯度达95%以上的rhOP-1蛋白,透析复性,加入L-精氨酸作为复性助溶剂,制备成冻干粉末,以不加助溶剂的rhOP-1为对照。将制备的冻干粉末复溶后,观察rhOP-1溶解性,并检测其活性。结果复性液中加入L-精氨酸的复性rhOP-1蛋白浓度为0.5mg/ml,回收率可达60%,冻干粉末复溶后,溶解性及rhOP-1活性良好。结论用此复性方法制备的rhOP-1溶液作为注射剂,有较好的应用前景。
Objective To prepare soluble recombinant human osteogenic protein-1 （rhOP-1） and explore the feasibility of preparation of rhOP-I injection. Methods Culture the recombinant E.coli for expression of rhOP-1 by fermentation, collect the bacteria, extract inclusion body and purify by SP-FF cation exchange chromatography. The rhOP-1 with a purity of more than 95% was reaturalized by dialysis, added with L-arginine as solubilizing agent and prepared into freeze-dried powder, using that without addition of L-arginine as control. The freeze-dried powder was reconstituted, then observed for solubility and determined for activity. Resuits The protein concentration and recovery of reaturalized rhOP-1 using L-arginine as solubilizing agent were 0. 5 mg/ ml and 60% respectively. The freeze-dried powder containing L-arginine showed good solubility and rhOP-1 activity after reconstitution. Conclusion The prepared rhOP-1 solution may be used as an injection of good prospect.
出处
《中国生物制品学杂志》
CAS
CSCD
2008年第7期605-607,共3页
Chinese Journal of Biologicals
基金
国家高技术研究发展计划(2003AA2Z3532)
关键词
重组人成骨蛋白-1
L-精氨酸
复性
注射剂
Recombinant human osteogenic protein-1
L-arginine
Renaturation
Injection
