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抗 Tityus caripitensis 毒液的 IgY 抗体:纯化和中和效果。

IgY antibodies anti-Tityus caripitensis venom: purification and neutralization efficacy.

机构信息

Centro de Biociencias y Medicina Molecular, Instituto de Estudios Avanzados-IDEA, Apartado 17606, Caracas 1015-A, Venezuela.

出版信息

Toxicon. 2013 Nov;74:208-14. doi: 10.1016/j.toxicon.2013.08.058. Epub 2013 Aug 29.

Abstract

Tityus caripitensis is responsible for most of scorpion stings related to human incidents in Northeastern Venezuela. The only treatment for scorpion envenomation is immunotherapy based on administration of scorpion anti-venom produced in horses. Avian antibodies (IgY) isolated from chicken egg yolks represent a new alternative to be applied as anti-venom therapy. For this reason, we produced IgY antibodies against T. caripitensis scorpion venom and evaluated its neutralizing capacity. The anti-scorpion venom antibodies were purified by precipitation techniques with polyethylene glycol and evaluated by Multiple Antigen Blot Assay (MABA), an indirect ELISA, and Western blot assays. The lethality neutralization was evaluated by preincubating the venom together with the anti-venom prior to testing. The IgY immunoreactivity was demonstrated by a dose-dependent inhibition in Western blot assays where antibodies pre-absorbed with the venom did not recognize the venom proteins from T. caripitensis. The anti-venom was effective in neutralizing 2LD50 doses of T. caripitensis venom (97.8 mg of IgY neutralized 1 mg of T. caripitensis venom). Our results support the future use of avian anti-scorpion venom as an alternative to conventional equine anti-venom therapy in our country.

摘要

特立尼达卡罗皮坦斯蝎是委内瑞拉东北部与人类有关的蝎子蜇伤事件的主要罪魁祸首。蝎子蜇伤的唯一治疗方法是基于在马身上产生的蝎子抗毒液的免疫疗法。从鸡蛋黄中分离出的禽抗体(IgY)代表了一种新的替代方法,可作为抗毒液疗法应用。出于这个原因,我们生产了针对特立尼达卡罗皮坦斯蝎毒液的 IgY 抗体,并评估了其中和能力。使用聚乙二醇沉淀技术对抗蝎毒液抗体进行纯化,并通过多抗原印迹分析(MABA)、间接 ELISA 和 Western blot 分析进行评估。通过在测试前将毒液与抗毒液预孵育来评估致死性中和。通过 Western blot 分析证明了 IgY 的免疫反应性,其中预先用毒液吸收的抗体无法识别来自特立尼达卡罗皮坦斯蝎的毒液蛋白。该抗毒液能够有效中和 2LD50 剂量的特立尼达卡罗皮坦斯蝎毒液(97.8mg 的 IgY 中和 1mg 的特立尼达卡罗皮坦斯蝎毒液)。我们的结果支持在我国未来使用禽抗蝎毒液作为传统马抗毒液治疗的替代方法。

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