Application of postsynaptic neurotoxin, cardiotoxin, cytotoxin, phospholipase A2 and crude venom of cobra in resisting virus infection
A cobra family, neurotoxin technology, applied in antiviral agents, medical preparations containing active ingredients, enzymes, etc.
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Embodiment 1
[0151] Example 1. Separation and purification of Chinese cobra crude venom to prepare postsynaptic neurotoxin, cytotoxin, cardiotoxin and phospholipase A2
[0152] Carry out cation exchange by TSK CM-650 (M) column with crude venom of cobra sinensis, the method for separating various toxins comprises the following steps:
[0153] 1. Sample preparation-dissolve 1g of Chinese cobra venom in the ammonium acetate buffer of 25ml 0.025 mole pH6.0, centrifuge at low temperature, and get the supernatant;
[0154] II. Equilibrium - equilibrate the TSK CM-650(M) column with 0.025 mole ammonium acetate solution of pH 6.0;
[0155] III. Elution - After loading the sample, use 0.1-0.5 mole and 0.7-1.0 mole ammonium acetate buffer solution, pH 5.9, to carry out 2-chamber gradient elution, UV detection parameter: 280nm; elution flow rate: 48ml / h;
[0156] IV. Collect various toxin components according to the recorded spectrum, and 12 protein peaks are eluted from the collected solution;
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Embodiment 2
[0159] Example 2. The isolated Chinese cobra post-synaptic neurotoxin, cytotoxin, cardiotoxin, phospholipase A2, and Chinese cobra crude venom were used for antiviral activity experiments, that is, virus plaque reduction rate % experiments
[0160] Principle of viral plaque
[0161] On a single layer of cells covered with a thin layer of agar, after the virus infects a certain cell, due to the limitation of the solid medium, the released virus can only expand from the initially infected cell to the periphery, that is to say, the proliferated virus particles can only spread to neighboring cells. After several cycles of proliferation, a localized area of diseased cells forms, eventually forming a plaque similar to a phage plaque. A plaque is formed by the replicative spread of a virus particle that initially infected a cell and is therefore an accurate measure of virus infectivity.
[0162] Plaque reduction rate% is an indicator of the ability to inhibit virus infection, and...
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