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用于被动免疫的鸡卵黄中蝰蛇抗蛇毒血清的研制

Study on development of Vipera lebetina snake anti-venom in chicken egg yolk for passive immunization.

作者信息

Zolfagharian Hossein, Dounighi Naser Mohammadpour

机构信息

a Department of Human Vaccine and Serum ; Razi Vaccine and Serum Research Institute ; Karaj , Iran.

出版信息

Hum Vaccin Immunother. 2015;11(11):2734-9. doi: 10.4161/21645515.2014.985492. Epub 2015 Feb 20.

Abstract

Chicken egg yolk antibodies against Vipera lebetina venom were evaluated for their antivenom potential. White leghorn hens were immunized with detoxified V. lebetina venom (γ-irradiated venom). The detoxified venom (200 μg) was mixed with an equal volume of complete Freund's adjuvant and was injected intramuscularly into the hens. The antibodies showed high activity (1.6 LD50/mL) in egg yolks after 12 d of venom injection. The eggs were collected after 12 days, and the egg yolks were removed and washed with purified water to remove any contamination with egg whites. The purification was performed using a method described by Maya Devi et al., followed by gel filtration (Sephadex G-50). The purity and molecular weight of antivenom antibodies (IgY) were determined using electrophoresis, and the molecular weight was found to be approximately 185 kDa. The potency of IgY was 6 LD50/mL (mice), i.e., 1 mL of IgY could neutralize 43.8 μg of standard V. lebetina venom). Our results showed that chicken egg yolk antibodies were effective in neutralizing the lethality and several pharmacological effects of V. lebetina venom and could be used for developing effective antivenom.

摘要

对针对埃及蝰蛇毒液的鸡卵黄抗体的抗蛇毒潜力进行了评估。用解毒的埃及蝰蛇毒液(γ射线辐照毒液)免疫白来亨鸡。将解毒毒液(200μg)与等体积的完全弗氏佐剂混合,肌肉注射到母鸡体内。注射毒液12天后,抗体在卵黄中显示出高活性(1.6 LD50/mL)。12天后收集鸡蛋,去除卵黄并用纯净水洗涤以去除蛋清的任何污染。使用Maya Devi等人描述的方法进行纯化,随后进行凝胶过滤(葡聚糖G-50)。使用电泳测定抗蛇毒抗体(IgY)的纯度和分子量,发现分子量约为185 kDa。IgY的效价为6 LD50/mL(小鼠),即1 mL IgY可以中和43.8μg标准埃及蝰蛇毒液。我们的结果表明,鸡卵黄抗体在中和埃及蝰蛇毒液的致死性和几种药理作用方面是有效的,可用于开发有效的抗蛇毒血清。

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