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

A cytotoxin and cobra venom technology, which is applied in the preparation methods of peptides, chemical instruments and methods, and medical preparations containing active ingredients, etc.

Inactive Publication Date: 2017-08-29
GUANGXI MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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

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Embodiment Construction

[0029] 1. Materials and methods

[0030] 1.1 Instruments and reagents

[0031] Electronic analytical balance (Sartotius, UK), start chromatography system (GE Company), L18-4584-01 system (PharmaciaBiotech Company of the United States), YC-2 chromatography experiment freezer (Beijing Boyikang Experimental Instrument Co., Ltd.), LGJ-18 freeze dryer (Beijing Songyuan Huaxing Science and Technology Development Company), microplate reader (Biotek Company, USA), cell culture box (Thermo Company), rat hepatic stellate cell HSC-T6 (Kunming Cell Bank, Type Culture Collection Committee, Chinese Academy of Sciences), DMEM medium (Gibco Company ), fetal bovine serum (Gibco company), CCK-8 reagent (Japanese colleagues), PE AnnexinV Apoptosis Detection Kit I (BD company in the United States), Guangxi cobra venom freeze-dried powder (provided by the Snake Venom Institute of Guangxi Medical University, cobras are reptiles) Lepidoptera suborder Cobra family Cobra genus, Zhoushan Cobra, Lat...

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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 ...

Claims

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Application Information

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IPC IPC(8): C07K14/46C07K1/18C07K1/16C07K1/14A61K38/17A61P1/16
CPCA61K38/1703C07K14/46
Inventor 张学荣孙林罗小玲廖明班建东蔡凤桃陈缨农君王秀南余雷
Owner GUANGXI MEDICAL UNIVERSITY
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