Lyophilized viper antivenin and preparation method thereof

Abstract

The invention discloses a lyophilized viper antivenin and a preparation method thereof, belonging to biochemical products, more particularly relating to an antivenin and a preparation technology thereof. The mass ratio of the antivenin and viper venom is 15:1, which can specifically neutralize the viper venom injected to mouse; as determined by immunodiffusion, the immunoprecipitation line appears in case that the ratio of viper antivenin F(ab')2 in lyophilized form to viper venom is 8:1; and other detected items conform to the quality standard of antivenin in Chinese Pharmacopoeia 2010. The detection of Phenyl-Sepharose (low-sub) FF column chromatography result shows that: activity is centralized at eluting peak 1, micromolecule impurity proteins are centralized at penetration peal, eluting peak 2 and eluting peak 3. According to the technology of the invention, immune blood plasma is resulted from viper venom immune horse, IgG is prepared by salting out the immune blood plasma, and F(ab')2 active fragment is obtained after the IgG is subject to enzymolysis and purification by a hydrophobic column. The lyophilized viper antivenin has strong specificity, high potency and more than 85% of the purity of antivenin F(ab')2 in lyophilized form.

Description

technical field [0001] The invention belongs to biomedical products, in particular to antivenom and its preparation process. Background technique [0002] Vipers (Vipera russelli siamensi) belong to the family Viperidae, and are highly venomous snakes that mainly contain blood circulation toxins. Vipers in my country are mainly distributed in Guangxi, Guangdong, Fujian and Taiwan. Vipers are active day and night, curled up into groups, move slowly, and have a ferocious snake nature. Viper venom mainly contains blood circulation poison, and the toxic components are mainly coagulation toxins, as well as hemolytic toxins, fibrinolytic toxins, anticoagulant components, platelet accumulation and denaturation components, and hemorrhagic toxins. After a person is bitten, the onset is acute, the symptoms are severe, the wound is swollen and painful, bleeding, causing extensive coagulation in the blood vessel, DIC dangerous symptoms, the patient may cause anemia and jaundice, renal...

AI Technical Summary

Problems solved by technology

However, Viper rotundus is a subspecies of Viperidae snakes, and the antigen specificity of different subspecies is quite different. F(ab′) produced by a single snake venom immune plasma 2 It can only be specifically combined with the corresponding antigenic determinant. In addition, Viper has a large amount of detoxification at one time and strong toxicity, so its limitation is obvious; while the concentrated stock solution uses chloroform as a solvent and has a large content (1.4-0.6%) and alum flocculation precipitation Proteins have potential damage to the human body; on the other hand, not only the finished product obtained by serum hydrolysis and salting out has low purity (F(ab′) 2 ≥60%) lead to low antibody titers, and because the antiserum contains more large molecular weight IgG and Fc fragments that activate complement, it becomes the most important factor in clinical antiserum allergic reactions except individual differences. Therefore, if you want to be more thorough It is far from enough to remove these allergenic ingredients only by salting out in the preparation

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