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Background <p>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<sub>1</sub> 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).</p> Objective <p>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.</p> Methods <p>We performed polymerase chain reaction (PCR) analysis to confirm the integration of EL and HV genes in the F<sub>1</sub> 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.</p> Results <p>A PCR analysis confirmed the successful integration of both EL and HV mAb genes in the F<sub>1</sub> progeny. Additionally, (RT)-PCR and immunoblotting validated the expression of these transgenes, with EL and HV mAbs purified from the F<sub>1</sub> 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.</p> Conclusion <p>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.</p>

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Plant cross-fertilization for production of dual-specific antibodies targeting both Ebola virus–like particles and HER2 protein in F1 plants

  • 期刊论文

  • 发表日期：

    2025年1月23日

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

摘要

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 F1 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 F1 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 F1 progeny. Additionally, (RT)-PCR and immunoblotting validated the expression of these transgenes, with EL and HV mAbs purified from the F1 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.