摘要
目的对重组人骨唾液酸蛋白(rhBSP)进行稳定性及成骨活性研究,为进一步研究rhBSP功能活性,揭示相关病理机制奠定基础。方法Western blot检测rhBSP抗原性;SDS-PAGE蛋白电泳及毛细管电泳分析rhBSP的降解稳定性;碱性磷酸酶(ALP)试剂盒检测rhBSP对小鼠胚胎成骨细胞株(MC3T3-E1)ALP活性的影响。结果rhBSP能与BSP单抗稳定结合;rhBSP对温度敏感,在4和25℃极易降解,而在-20℃和冻干保存则相对稳定;rhBSP能提高MC3T3-E1细胞的ALP活性。结论rhBSP的稳定性主要受温度的影响,-20℃保存相对稳定;rhBSP可以促进MC3T3-E1细胞ALP的表达。
Purpose To analyze recombinant human bone sialoprotein （rhBSP） stability and ability of promoting alkaline phosphatase （ALP） activity of mouse osteoblastic cell line embryol （MC3T3-E1） so as to provide a basis for the deep research on biology and pathology of rhBSP. Methods The antigenicity of rhBSP was detected by western blot, the stability of degradation was analyzed by SDS-PAGE and capillary electrophoresis, the effect of rhBSP on ALP activity of MC3T3-E1 was examined by using ALP assay kit. Results rhBSP can combine with its monoclonal antibody stably. Stability analysis showed that the rhBSP was sensitive to temperature and easy to degrade at 4 ℃ and 25 ℃. However, it was relatively stable at - 20 ℃ or lyophilized. The rhBSP can promote ALP activity of MC3T3-E1. Conclusion Stability of rhBSP was mainly influenced by temperature, relatively stable at - 20 ℃. The rhBSP can induce the expression of ALP in MC3T3-E1.
出处
《中国生化药物杂志》
CAS
CSCD
北大核心
2009年第4期247-250,共4页
Chinese Journal of Biochemical Pharmaceutics
基金
广东省自然科学基金研究团队项目资助(编号:20023001)
广东省科技计划项目资助(编号:2005B34001008)
