两种不同产地哥伦比亚Bothrops asper实验单价抗蛇毒血清的中和作用比较

K. Sarmiento, IvonneTorres, C. Ríos, Julian Salazar, Andrea Baracaldo, Jorge Zambrano, German Ramírez-Forero, Diego Hernández, Luisa Pérez, Diana Castaño, H. Díez
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引用次数: 2

摘要

背景和目的:哥伦比亚不生产抗蛇毒血清,本研究的目的是比较来自大西洋和安第斯地区的Bothrops asper毒液与2种实验性单价抗蛇毒血清的中和作用。材料与方法:采用280 nm吸光度法对蛇毒及抗蛇毒血清蛋白进行定量分析,并采用SDS-PAGE进行结构表征。采用反相高效液相色谱法测定血清蛋白谱。使用Prism-statMate计算毒液的LD50和抗蛇毒血清的ED50。用安第斯蛇毒对马进行了为期3个月的免疫接种。用辛酸制备完整IgG抗蛇毒血清,用胃蛋白酶制备抗蛇毒血清F(ab ')2。两种抗蛇毒血清都用安第斯蛇毒和大西洋蛇毒进行了测试。电泳测定抗蛇毒血清的重量。结果:安第斯毒蛋白含量为11.5 mg/mL,大西洋毒蛋白含量为13.4 mg/mL。SDS-PAGE显示两种毒液的条带均为15 ~ 20 kDa。对安第斯毒液的LD50为2.8 mg/kg,对大西洋毒液的LD50为2.3 mg/kg。RT-HPLC显示两种毒液在前40分钟的蛋白质含量相似。IgG蛋白为9.8 mg/mL, F(ab’)2蛋白为11.2 mg/mL。SDS-PAGE显示IgG的条带为130 ~ 200 kDa, F(ab’)2的条带为94 ~ 150 kDa。IgG和F(ab’)2对大西洋毒液的ED50分别为1.4 mg/mL和1.75 mg/mL。用IgG和F(ab’)2对安第斯毒的ED50分别为1.6 mg/mL和1.9 mg/mL。结论:两种抗蛇毒血清均能有效中和安第斯蛇毒和大西洋蛇毒,且无显著性差异。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of Neutralization of Two Experimental Monovalent Antivenoms of Colombia's Bothrops asper from Different Localities
Background and Objectives: Colombia does not produce fabotherapic antivenoms, the aim of current study was comparison of the neutralization of Bothrops asper venom from Atlantic and Andean region with 2 experimental monovalent antivenoms. Materials and Methods: The protein of the venom and antivenoms was quantified by absorbance at 280 nm and characterized by SDS-PAGE. The protein profile venoms were performed by RT-HPLC. It was calculated LD50 for the venoms and ED50 for the antivenoms, using Prism-statMate. An immunization scheme of 3 months was conducted in equine using Andean venom. The complete IgG antivenom was produced with caprylic acid, while the fabotherapic F(ab’)2 antivenom with pepsin. Both antivenoms were tested with Andean venom and Atlantic venom. Electrophoresis was conducted to verify the antivenoms by weight. Results: The protein was 11.5 mg/mL to Andean venom and 13.4 mg/mL to Atlantic venom. The SDS-PAGE showed bands of 15-20 kDa for both venoms. The LD50 to Andean venom was 2.8 and 2.3 mg/kg to Atlantic venom. The RT-HPLC showed similar protein fractions at first 40 min in both venoms. The protein for IgG was 9.8 mg/mL, while for F(ab’)2 it was 11.2 mg/mL. The SDS-PAGE showed bands of 130-200 kDa for IgG whyle of 94-150 kDa for F(ab’)2. The ED50 with Atlantic venom was 1.4 mg/mL using IgG and 1.75 mg/mL using F(ab’)2. The ED50 with Andean venom was 1.6 mg/mL using IgG and 1.9 mg/mL using F(ab’)2. Conclusion: Both antivenoms were efficient to neutralize the venom of Andean and Atlantic venom, without significant differences.
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