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Expression of anti human IL-4 and IL-6 scFvs in transgenic tobacco plants

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Abstract

The two murine single-chain Fv (scFv) genes against human interleukin IL-4 and IL-6 cytokines were cloned in a plant expression vector (pGEJAE1) and mobilized to Agrobacterium tumefaciens. Tobacco leaf discs were co-cultured with Agrobacterium and transferred to selective media for regeneration. The tobacco in vitro plants produced scFvs against human IL-4 and IL-6. Only 8% of transformed plants expressing anti-IL-4 scFv were obtained versus 76% of transformed plants expressing anti-IL-6 scFv. In addition, some plants producing anti-IL-4 and anti-IL-6 scFvs aged more rapidly in in vitro conditions and in greenhouse pots than did control plants. Western blot analysis showed that the transformed Nicotiana tabacum plants contained proteins with an apparent molecular mass on electrophoresis of ca. 32 kDa, corresponding to the predicted size of the scFvs. As entire plant root seemed to accumulate more scFv than did leaves, we decided to continue working with isolated roots. Anti-IL-6 scFvs were detected in cultivated roots and their culture media. Functional anti-IL-6 scFv accounted for 0.16–0.18% of total soluble proteins. The affinity of the anti-IL-6 scFv produced in plants and measured by Biacore was similar to that of scFv produced in Escherichia coli. The high levels of antibody accumulation in isolated roots and secretion into the medium demonstrate the potential for producing recombinant protein in bioreactor systems.

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Authors and Affiliations

  1. Molecular Biology Unit, Institut Pasteur, Avenue Pasteur, 13164, Teheran, Iran

    P. Ehsani & A. Jafari

  2. Laboratoire Ingénierie des Anticorps, Institut Pasteur, 25–28 rue du Docteur Roux, 75724, Paris Cedex 15, France

    A. Meunier, F. Nato, A. Nato & P. Lafaye

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  1. P. Ehsani
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  2. A. Meunier
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  3. F. Nato
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  4. A. Jafari
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  5. A. Nato
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  6. P. Lafaye
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Correspondence to P. Lafaye.

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these authors contributed equally to this work

these authors contributed equally to this work

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Ehsani, P., Meunier, A., Nato, F. et al. Expression of anti human IL-4 and IL-6 scFvs in transgenic tobacco plants. Plant Mol Biol 52, 17–29 (2003). https://doi.org/10.1023/A:1023902407855

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