Purification preparation method and application of cobra venom CTX (cytotoxin)-4N

Abstract

The invention discloses a purification preparation method of cobra venom CTX (cytotoxin)-4N. According to the method, a DEAE-SepharoseCL-6B anion exchange column is firstly used; then, a SephadexG-50 gel chromatographic column and a Macro-prep High S cation exchange column are used for respective separation; after separation in each time, the peak, with the highest activity, of the cobra venom CTX-4N is selected as a sample of the subsequent separation. Research shows after the method is used, CTX-4N with electrophoresis purity can be obtained; the purity is high; the activity is high. HSC-T6 proliferation inhibition and apoptosis experiments prove that the CTX-4N has a killing effect on HSC-T6 cells; the condition that the CTX-4N has the anti-hepatic fibrosis effect in the hepatic fibrosis development process is prompted; important medicinal values are realized.

Description

technical field [0001] The invention belongs to the field of cobra venom cytotoxin, and in particular relates to a purification preparation method and application of cobra venom cytotoxin-4N (CTX-4N). Background technique [0002] Cobra venom is secreted by the venom glands of cobras. Its chemical composition is complex, containing a variety of proteins, polypeptides, enzymes and other small molecules, and has a wide range of biological activities. Cobra cytotoxin (Cytotoxin, CTX), also known as cobra cardiotoxin (Cardiototxin), is one of the important active substances of cobra venom, accounting for about 25% to 40% of snake venom protein. Polarization, antibacterial and other biological activities, and induce apoptosis of a variety of tumor cells, the biological effects of components contained in snake venoms of different snake species in different regions are not the same. [0003] Isolation and purification of CTX from cobra is a necessary prerequisite for the study of ...

AI Technical Summary

Problems solved by technology

In terms of separation and purification of CTX, domestic and foreign scholars mostly adopt the method of combining cation exchange column chromatography and gel filtration to carry out separation and purification, and found CTX-I, CTX-II, CTX-III, CTX-IV (SEQ. ID.No.2), CTX-V and other cytotoxins. However, in this method, due to the many factors affecting the column efficiency, and it is difficult to control, the repeatability is poor, and the products obtained after separation and purification often contain other protein substances. , which will affect the study of the biological activity of cytotoxins

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