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Purified the Hemorrhagic Toxin of Agkistrodon acutus Venom and Its Antibody Applied in Identification Different Venoms |
ZHENG Ying, ZHANG Yi, YE Feng-ping, Nie Xue-feng, ZHANG Zhi-xiao, Qiu Wei, FAN Quan-shui |
Military Medical Institute,CDC of PLA Chengdu Military Area Command,Kunming 650118,China |
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Abstract Aim: To purify the hemorrhagic toxin (HR) of Agkistrodon acutus venom, prepare its special antibody and establish related ELISA method for detection of different snake venoms. Methods: HR was obtained from Agkistrodon acutus venom by several column chromatographs and used as antigen to immunized rabbits. IgG antibody was isolated and purified from immunized rabbit plasma by ammonium sulfate precipitate and affinity chromatography, labeled with HRP. The five venoms from Viperidae were identified by developed ELISA using purified specific igG antibody. Results: The purity of purified HR was up to 99.49 %, ELISA analysis suggested that the specific HRP-IgG antibody can be fast and exactly identified venom of Agkistrodon acutus from other viperidae venoms, and its sensitivity was 10ng/ml, the rate of cross reaction was <15%. Conclusion: The developed method of purified HR antibody sandwich ELISA can be quickly and accurately identified venom of Agkistrodon acutus from other kinds of viperidae venoms.
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Received: 26 November 2012
Published: 25 March 2013
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