Separation and purification of the anti-clotting protein component from venom of Agkistrodon and its acute toxicity testing
- VernacularTitle:皖南尖吻蝮蛇毒抗凝蛋白组分的分离纯化及其急性毒性实验研究
- Author:
Jianguo HUAI
;
Jing RUI
- Publication Type:Journal Article
- Keywords:
snake venom;
anticoagulation;
acute toxicity;
LD50
- From:
Chinese Journal of Clinical Pharmacology and Therapeutics
2000;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To separate and purify a new anti-clotting protein component(ACPF-7221)from the venom of Agkistrodon. The acute toxicity experiment of ACPF-7221 and the death rate were studied and the LD50 and 95% confidence interval were determined in mice. METHODS: The venom of Agkistrodon was separated and purified using ion-exchange chromatography of DEAE-Sepharose Fast Flow, molecular sieve filtration through Sephadex G75, SP-Sepharose Fast Flow and molecular sieve filtration through Sephadex G50, the anticoagulation components were recalled using activated partial thromboplastin time (APTT) activity assay. ACPF-7221 was given to mice by intravenous injection,and the dosage was determined according to the body weight of mice. The LD50 and 95% confidence were calculated by the death outcome. RESULTS: ACPF-7221 was separated and purified,which was a trimer consisted of three kinds of protein and their relative molecular mass were 25000, 30000 and 50000,and the protein level was 65%. The LD50 of ACPF-7221 was 7.848 mg/kg, and its 95% confident interval was 7.353-8.375 mg/kg. CONCLUSION: A new anti-clotting protein component ACPF-7221 was extracted from the venom of Agkistrodon by ion-exchange chromatography,which is a trimer consisted of three kinds of protein. The critical orga of acute toxicity is lung, empsyxis asphyxia deduces death.
