Recombinant antibodies against Iranian cobra venom as a new emerging therapy by phage display technology.

Ali Nazari, Maedeh Samianifard, Hadi Rabie, Abbas Zare Mirakabadi
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引用次数: 5

Abstract

Background: The production of antivenom from immunized animals is an established treatment for snakebites; however, antibody phage display technology may have the capacity to delivery results more quickly and with a better match to local need. Naja oxiana, the Iranian cobra, is a medically important species, responsible for a significant number of deaths annually. This study was designed as proof of principle to determine whether recombinant antibodies with the capacity to neutralize cobra venom could be isolated by phage display.

Methods: Toxic fractions from cobra venom were prepared by chromatography and used as targets in phage display to isolate recombinant antibodies from a human scFv library. Candidate antibodies were expressed in E. coli HB2151 and purified by IMAC chromatography. The selected clones were analyzed in in vivo and in vitro experiments.

Results: Venom toxicity was contained in two fractions. Around a hundred phage clones were isolated against each fraction, those showing the best promise were G12F3 and G1F4. While all chosen clones showed low but detectable neutralizing effect against Naja oxiana venom, clone G12F3 could inhibit PLA2 activity.

Conclusion: Therefore, phage display is believed to have a good potential as an approach to the development of snake antivenom.

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利用噬菌体展示技术制备伊朗眼镜蛇毒液重组抗体。
背景:从免疫动物中生产抗蛇毒血清是治疗蛇咬伤的一种成熟方法;然而,抗体噬菌体展示技术可能有能力更快地提供结果,并更好地满足当地的需求。伊朗眼镜蛇Naja xiana是医学上重要的物种,每年造成大量死亡。本研究旨在验证噬菌体展示法能否分离出具有中和眼镜蛇毒液能力的重组抗体。方法:采用层析法制备眼镜蛇毒液毒性组分,并将其作为噬菌体展示的靶点,从人单链病毒文库中分离重组抗体。候选抗体在大肠杆菌HB2151中表达,经IMAC层析纯化。对所选克隆进行体内和体外实验分析。结果:毒液毒性分为两部分。从每个片段中分离出大约100个噬菌体克隆,其中最有希望的是G12F3和g14f4。克隆G12F3能抑制PLA2活性,而克隆G12F3能抑制PLA2活性。结论:噬菌体展示技术在开发抗蛇毒血清中具有良好的应用前景。
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来源期刊
CiteScore
4.80
自引率
8.30%
发文量
39
审稿时长
6-12 weeks
期刊介绍: Journal of Venomous Animals and Toxins including Tropical Diseases (JVATiTD) is a non-commercial academic open access publication dedicated to research on all aspects of toxinology, venomous animals and tropical diseases. Its interdisciplinary content includes original scientific articles covering research on toxins derived from animals, plants and microorganisms. Topics of interest include, but are not limited to:systematics and morphology of venomous animals;physiology, biochemistry, pharmacology and immunology of toxins;epidemiology, clinical aspects and treatment of envenoming by different animals, plants and microorganisms;development and evaluation of antivenoms and toxin-derivative products;epidemiology, clinical aspects and treatment of tropical diseases (caused by virus, bacteria, algae, fungi and parasites) including the neglected tropical diseases (NTDs) defined by the World Health Organization.
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