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通过植物杂交受精在F植物中生产同时靶向埃博拉病毒样颗粒和HER2蛋白的双特异性抗体。

Plant cross-fertilization for production of dual-specific antibodies targeting both Ebola virus-like particles and HER2 protein in F plants.

作者信息

Lee Daehwan, Hwang Hyunjoo, Kim Yerin, Hwang Yejin, Youk Keunbeom, Hinterdorfer Peter, Kim Mikyung, Ko Kisung

机构信息

Department of Medicine, BioSystems Design Lab, College of Medicine, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul, 06974, Korea.

Department of Applied Experimental Biophysics, Johannes Kepler University Linz, Linz, 4040, Austria.

出版信息

Genes Genomics. 2025 Apr;47(4):425-433. doi: 10.1007/s13258-025-01616-z. Epub 2025 Jan 23.

Abstract

BACKGROUND

This study explores the cross-fertilization of transgenic tobacco plants to produce dual-specific monoclonal antibodies (mAbs) targeting Ebola virus-like particles and HER2 proteins. We generated F plants by hybridizing individual transgenic lines expressing the anti-HER2 breast cancer VHH mAb (HV) and the H-13F6 human anti-Ebola large single chain mAb (EL).

OBJECTIVE

Hybridizing transgenic plants to express dual-antibodies between different structures VHH and LSCK indicate the potential of transgenic plants as a cost-effective and scalable production system for dual targeting mAbs.

METHODS

We performed polymerase chain reaction (PCR) analysis to confirm the integration of EL and HV genes in the F progeny. The reverse-transcription (RT)-PCR and immunoblotting were performed to confirm the expression of transgenes. Indirect enzyme-linked immunosorbent assay was conducted to confirm the functionality of purified EL and HV mAb.

RESULTS

A PCR analysis confirmed the successful integration of both EL and HV mAb genes in the F progeny. Additionally, (RT)-PCR and immunoblotting validated the expression of these transgenes, with EL and HV mAbs purified from the F plants. Indirect enzyme-linked immunosorbent assay (ELISA) demonstrated that EL × HV mAb proteins maintained binding activity to Ebola virus-specific antigens, comparable to that of the EL mAb protein, while also exhibiting binding activity against HER2 proteins similar to that of the HV mAb.

CONCLUSION

This study indicates the potential for transgenic plants to produce dually targeting mAbs, suggesting a promising application in enabling the co-expression of antibodies targeting two different diseases in a single plant.

摘要

背景

本研究探索转基因烟草植物的杂交,以产生靶向埃博拉病毒样颗粒和HER2蛋白的双特异性单克隆抗体(mAb)。我们通过将表达抗HER2乳腺癌VHH单克隆抗体(HV)的单个转基因品系与H-13F6人抗埃博拉大的单链单克隆抗体(EL)杂交,培育出了F植株。

目的

使转基因植物杂交以表达不同结构VHH和LSCK之间的双抗体,表明转基因植物作为双靶向单克隆抗体的经济高效且可扩展生产系统的潜力。

方法

我们进行聚合酶链反应(PCR)分析,以确认EL和HV基因在F后代中的整合。进行逆转录(RT)-PCR和免疫印迹以确认转基因的表达。进行间接酶联免疫吸附测定,以确认纯化的EL和HV单克隆抗体的功能。

结果

PCR分析证实EL和HV单克隆抗体基因均成功整合到F后代中。此外,(RT)-PCR和免疫印迹验证了这些转基因的表达,从F植株中纯化出了EL和HV单克隆抗体。间接酶联免疫吸附测定(ELISA)表明,EL×HV单克隆抗体蛋白对埃博拉病毒特异性抗原保持结合活性,与EL单克隆抗体蛋白相当,同时对HER2蛋白也表现出与HV单克隆抗体相似的结合活性。

结论

本研究表明转基因植物具有产生双靶向单克隆抗体的潜力,提示在使单株植物共表达靶向两种不同疾病的抗体方面具有广阔的应用前景。

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