舟山眼镜蛇毒CTX1层析分离的流速优化

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...

Flow velocitiy optimization of isolation methods for component CTX1 of Naja naja atra venom

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.