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通过噬菌体展示技术制备抗伊朗眼镜蛇毒重组抗体作为一种新兴疗法

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

作者信息

Nazari Ali, Samianifard Maedeh, Rabie Hadi, Mirakabadi Abbas Zare

机构信息

Department of Biochemistry and Proteomics, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

Department of Venomous Animals, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

出版信息

J Venom Anim Toxins Incl Trop Dis. 2020 Jun 19;26:e20190099. doi: 10.1590/1678-9199-JVATITD-2019-0099.

DOI:10.1590/1678-9199-JVATITD-2019-0099
PMID:32695146
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7346683/
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. , 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 HB2151 and purified by IMAC chromatography. The selected clones were analyzed in and 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 venom, clone G12F3 could inhibit PLA activity.

CONCLUSION

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

摘要

背景

用免疫动物生产抗蛇毒血清是治疗蛇咬伤的既定方法；然而，抗体噬菌体展示技术可能有能力更快地产生结果，并且与当地需求更匹配。伊朗眼镜蛇是一种具有重要医学意义的物种，每年导致大量死亡。本研究旨在作为原理验证，以确定是否可以通过噬菌体展示分离出具有中和眼镜蛇毒液能力的重组抗体。

方法

通过色谱法制备眼镜蛇毒液的有毒部分，并将其用作噬菌体展示的靶标，以从人单链抗体库中分离重组抗体。候选抗体在HB2151中表达，并通过IMAC色谱法纯化。对所选克隆进行了[具体实验名称1]和[具体实验名称2]实验分析。

结果

毒液毒性存在于两个部分中。针对每个部分分离出了约一百个噬菌体克隆，其中最有前景的是G12F3和G1F4。虽然所有选定的克隆对[毒液名称]毒液显示出低但可检测到的中和作用，但克隆G12F3可以抑制PLA活性。

结论

因此，噬菌体展示被认为作为一种开发蛇抗蛇毒血清的方法具有良好的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/04d68c22deb0/1678-9199-jvatitd-26-e20190099-gf5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/b27dbe0218cd/1678-9199-jvatitd-26-e20190099-gf1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/f10540d13fa6/1678-9199-jvatitd-26-e20190099-gf2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/eb609547ae64/1678-9199-jvatitd-26-e20190099-gf3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/badd06e73071/1678-9199-jvatitd-26-e20190099-gf4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/04d68c22deb0/1678-9199-jvatitd-26-e20190099-gf5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/b27dbe0218cd/1678-9199-jvatitd-26-e20190099-gf1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/f10540d13fa6/1678-9199-jvatitd-26-e20190099-gf2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/eb609547ae64/1678-9199-jvatitd-26-e20190099-gf3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/badd06e73071/1678-9199-jvatitd-26-e20190099-gf4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fca1/7346683/04d68c22deb0/1678-9199-jvatitd-26-e20190099-gf5.jpg

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