摘要
目的比较3种流速的CM-Sepharose FF阳离子交换层析柱分离舟山眼镜蛇(Naja naja atra)蛇毒(snake venom,SV)的柱效,为SV的分离纯化提供实验依据。方法 (1)采用3种流速CM-Sepharose FF阳离子交换层析柱分离舟山眼镜蛇蛇毒;(2)反向高效液相法分析各组分的纯度及内标法与CTX1标准品比较指纹图谱。结果经过阳离子交换层析,可从蛇毒中获得7个组分(组分1～7)。当流速为1.4 ml/min和1.8ml/min时,得到的7个组分中,组分SV1和SV2,组分SV3和SV4,组分SV5、SV6和SV7组分之间分离效果较差;当流速为1.6 ml/min时,各组分的分离效果理想。HPLC分析显示,组分SV5、组分SV6都是单一成分,纯度分别为95.8%、93.6%;CTX1内标法显示,组分SV6与CTX1指纹图谱吻合。结论 CM-Sepharose FF用于蛇毒分离可获得较高纯度的单一有效成分,流速为1.6 ml/min分离效果佳。
Objective Comparing the column efficiency of three flow velocities in CM-Sepha- rose FF ion-exchange chromatography gel chromatography on isolating Naja naja atra snake venom,it is order to research and develop new method to purificate and characterizate SV. Methods （1）Isolation and purification of venom was done by successive chromatography CM- Sepharose Fast Flow in three flow velocities; （2）The characterizations of this protein were studied by HPLC. Results Seven components（SV1-SV7）,were obtained from Naja naja atra snake venom by CM-Sepharose FF chromatography column. When flow velocity changed from 1.4 ml/min to 1.6 ml/min to 1.8 ml/min,SV do not been separated completely at flow velocity 1.4 ml/min and 1.8 ml/min,while been separated completely at flow velocity 1.6 ml/ rain. HPLC analysis indicated SV5 and SV6 were sterling. The purity were 95.8% and 93.6%. HPLC internal standard method analysis indicated that SV6 was the same with CTX1 in fingerprint. Conclusion The parameters about flow velocity of 1.6 ml/min linear gradient were more optimum in isolation and purification of SV in this experiment.
出处
《蛇志》
2011年第2期96-101,共6页
Journal of Snake
基金
广州市科技局资助项目(No.穗科条字[2005]6号)
关键词
舟山眼镜蛇
蛇毒
纯化
鉴定
Naja naja atra
snake venom
purification
characterization
