Generation of chicken-based IgY polyclonal antibodies against Dendroaspis polylepis and preclinical evaluation of envenomation-neutralizing efficacy vis-à-vis selected commercial antivenoms

IF 3.6 Q2 TOXICOLOGY Toxicon: X Pub Date : 2024-06-28 DOI:10.1016/j.toxcx.2024.100201
Stephen Wilson Kpordze , Victor Atunga Mobegi , Gideon Mutie Kikuvi , Joseph Kangangi Gikunju , Courage Kosi Setsoafia Saba , Jackan Moshe , James Hungo Kimotho
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Abstract

The Black mamba, D. polylepis, is one of the many venomous snakes found in Kenya, and known to account for some snakebite incidents. The Kenyan Ministry of Health data reveals annual 15,000 snakebites occurrences. Also, 1 in 15 people in Kenya gets bitten by a snake, and tragically, 1 in 147 of these individuals die of snakebite yearly. Traditionally, antivenoms for treatment are produced from horse or sheep but have complicated and expensive production issues. Alternative production approaches, such as using IgY antibodies derived from chicken egg yolks, may overcome disadvantages with traditional antivenom manufacturing techniques. In this current study, D. polylepis specific IgY polyclonal antibodies were purified from the egg yolks of chickens immunized with D. polylepis venom. These antibodies were subsequently assessed for their in-vivo neutralizing capacity vis-à-vis commercial antivenoms, PANAF-Premium and VINS. The IgY antibodies were purified by ammonium sulfate precipitation and affinity-chromatography, with quality and specificity determined by SDS-PAGE and ELISA. The LD50 of D. polylepis was found to be 0.54 mg/kg in chicks, and 0.34 mg/kg in mice, respectively. Pool of extracted IgY yielded 2.8 mg/mL concentration. Purified IgY under non-reducing and reducing conditions on SDS-PAGE exhibited a single-protein band of about 183 kDa and two bands (67 kDa and 25 kDa), respectively. The minimum-edematogenic dose was 0.05 μg. Anti-D. polylepis IgY antibodies and two antivenoms demonstrated the capacity to neutralize the toxic activities of D. polylepis venom. This study confirms a successful IgY generation against Black mamba venom for the first time, and observed toxic effects of the venom as well as neutralizing capacity of antivenoms.

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产生鸡基 IgY 多克隆抗体抗多角雉(Dendroaspis polylepis),并对与选定的商业抗蛇毒血清相比的蛇毒中和效力进行临床前评估
黑曼巴蛇(D. polylepis)是肯尼亚发现的众多毒蛇之一,据悉也是造成一些蛇咬伤事件的原因。肯尼亚卫生部的数据显示,每年发生 15,000 起蛇咬事件。此外,肯尼亚每 15 人中就有 1 人被蛇咬伤,而不幸的是,其中每年每 147 人中就有 1 人死于蛇咬。传统上,用于治疗的抗蛇毒血清是从马或羊身上提取的,但生产过程复杂且成本高昂。替代生产方法,如使用从鸡蛋黄中提取的 IgY 抗体,可以克服传统抗蛇毒血清生产技术的缺点。在本研究中,从用多刺蛇毒免疫的鸡蛋黄中纯化出了多刺蛇特异性 IgY 多克隆抗体。随后对这些抗体与商业抗蛇毒血清 PANAF-Premium 和 VINS 的体内中和能力进行了评估。IgY 抗体是通过硫酸铵沉淀和亲和层析法纯化的,其质量和特异性由 SDS-PAGE 和 ELISA 测定。小鸡和小鼠的半数致死剂量分别为 0.54 毫克/千克和 0.34 毫克/千克。提取的 IgY 池浓度为 2.8 毫克/毫升。在非还原和还原条件下,纯化的 IgY 在 SDS-PAGE 上分别显示出约 183 kDa 的单蛋白条带和两条条带(67 kDa 和 25 kDa)。最小致畸剂量为 0.05 μg。抗多头蝙蝠 IgY 抗体和两种抗蛇毒血清证明了中和多头蝙蝠毒液毒性的能力。这项研究首次证实了针对黑曼巴毒液的IgY抗体的成功产生,并观察到了毒液的毒性作用以及抗蛇毒血清的中和能力。
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来源期刊
Toxicon: X
Toxicon: X Pharmacology, Toxicology and Pharmaceutics-Toxicology
CiteScore
6.50
自引率
0.00%
发文量
33
审稿时长
14 weeks
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