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抗台湾烙铁头蛇蛇毒的单链抗体片段。

Single Chain Antibody Fragment against Venom from the Snake Daboia russelii formosensis.

机构信息

Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan.

The Center of Translational Medicine, Taipei Medical University, Taipei 11031, Taiwan.

出版信息

Toxins (Basel). 2017 Oct 27;9(11):347. doi: 10.3390/toxins9110347.

DOI:10.3390/toxins9110347
PMID:29076991
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5705962/
Abstract

Russell's vipers containing hemotoxic and neurotoxic venom commonly cause snake envenomation. Horse-derived antivenom is a specific antidote, but its production is expensive and has side effects. Developing a cost-effective and more tolerable therapeutic strategy is favorable. In this study, using glutaraldehyde-attenuated (DRF) venom proteins to immunize chickens, polyclonal yolk-immunoglobulin (IgY) antibodies were generated and showed a specific binding affinity. Phage display technology was used to generate two antibody libraries of single-chain variable fragments (scFvs) containing 3.4 × 10⁷ and 5.5 × 10⁷ transformants, respectively. Phage-based ELISA indicated that specific clones were enriched after bio-panning. The nucleotide sequences of scFv-expressing clones were analyzed and classified into six groups in the short linker and four groups in the long linker. These scFv antibodies specifically bound to DRF proteins, but not other venom proteins. Mass spectrometric data suggested that these scFv antibodies may recognize phospholipase A2 RV-4 or RV-7. In vivo studies showed that anti-DRF IgY exhibited complete protective effects and mixed scFv antibodies increased the survival rate and time of mice challenged with a lethal dose of DRF proteins. These antibodies can be potentially applied in a rapid diagnostic method or for treatment in the future.

摘要

罗素氏蝰蛇含有血液毒性和神经毒性毒液，通常会导致蛇咬伤。马源抗蛇毒血清是一种特效解毒剂，但生产成本高且有副作用。因此，开发一种经济有效且耐受性更好的治疗策略是有利的。在这项研究中，我们使用戊二醛减毒的（DRF）毒液蛋白对鸡进行免疫，产生了多克隆卵黄免疫球蛋白（IgY）抗体，并显示出特定的结合亲和力。噬菌体展示技术用于生成两个单链可变片段（scFv）抗体文库，分别包含 3.4×10⁷和 5.5×10⁷个转化体。噬菌体 ELISA 表明，生物淘选后特异性克隆得到了富集。对 scFv 表达克隆的核苷酸序列进行分析，并在短接头中分为六组，在长接头中分为四组。这些 scFv 抗体特异性结合 DRF 蛋白，但不与其他毒液蛋白结合。质谱数据表明，这些 scFv 抗体可能识别磷脂酶 A2 RV-4 或 RV-7。体内研究表明，抗-DRF IgY 表现出完全的保护作用，而混合 scFv 抗体则提高了接受致死剂量 DRF 蛋白攻击的小鼠的存活率和时间。这些抗体可能具有潜在的应用价值，可用于快速诊断方法或未来的治疗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/b4f7ea4e96d9/toxins-09-00347-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/48078a579586/toxins-09-00347-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/169802f67b5f/toxins-09-00347-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/41099b82f712/toxins-09-00347-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/98f736910038/toxins-09-00347-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/bcb9e6ccb77a/toxins-09-00347-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/8f5027996923/toxins-09-00347-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/68e9e5bb4874/toxins-09-00347-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/b4f7ea4e96d9/toxins-09-00347-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/48078a579586/toxins-09-00347-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/169802f67b5f/toxins-09-00347-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/41099b82f712/toxins-09-00347-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/98f736910038/toxins-09-00347-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/bcb9e6ccb77a/toxins-09-00347-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/8f5027996923/toxins-09-00347-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/68e9e5bb4874/toxins-09-00347-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea4b/5705962/b4f7ea4e96d9/toxins-09-00347-g008.jpg

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