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Transgenic Expression and Functional Characterization of Human Platelet Derived Growth Factor BB (hPDGF-BB) in Tobacco (Nicotiana tabacum L.)

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Abstract

Recombinant human platelet derived growth factor BB (rhPDGF-BB) is clinically approved for treating diabetic neuropathic ulcers. Plant-based expression systems offer less expensive ways of producing recombinant drugs, which do not require purification for clinical use. From this perspective, rhPDGF-BB is an ideal candidate for expression in plants as it can be applied topically. Here, we report a proof of concept study, in which rhPDGF-BB was expressed in tobacco plants, and its biological activity was tested in vitro. The mature human platelet derived growth factor BB (hPDGF-BB) gene was codon-optimized for tobacco and fused with ER targeting and retention signals, 5′ and 3′ UTRs of arc5-1 gene along with CaMV 35S promoter, and then, transferred by Agrobacterium-mediated transformation. Gene and protein expression of hPDGF-BB were confirmed by PCR and immunoblot studies. Bioactivity of hPDGF-BB expressed protein was determined by in vitro assays such as proliferation and migration in NIH3T3 cells. Our data reveals that total soluble proteins containing hPDGF-BB from transgenic plants showed a 4.5-fold increase in fibroblast proliferation compared to non-transgenic plants. Furthermore, plant-made rhPDGF-BB induced chemotaxis of treated cells and promoted wound healing in vitro. These results clearly demonstrate that functionally active rhPDGF-BB protein can be produced in plants and might have therapeutic benefits.

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Acknowledgments

We thank Dr. Carl Henrik Heldin, Ludwig Institute for Cancer Research, Uppsala, Sweden for providing the anti-hPDGF-BB antibody. The generous support of Mrs. Silke Brillof, University Center for Vascular Medicine, University Hospital Carl Gustav Carus, Technische Universität Dresden University of Dresden is gratefully acknowledged for assisting in cell culture experiments. This study was funded by SRM University. We thank Technische Universität Dresden for funding and support the experiments on cell culture work.

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Authors and Affiliations

  1. Genomics Laboratory, Department of Genetic Engineering, SRM University, Chennai, 603 203, Tamil Nadu, India

    Kanagasabapathy Deepa & Madasamy Parani

  2. Center for Vascular Medicine and Department of Medicine, Division of Angiology, University Hospital Carl Gustav Carus, Technische Universität Dresden, 01307, Dresden, Germany

    Roman N. Rodionov & Norbert Weiss

Authors
  1. Kanagasabapathy Deepa
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  2. Roman N. Rodionov
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  3. Norbert Weiss
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  4. Madasamy Parani
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Correspondence to Madasamy Parani.

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Deepa, K., Rodionov, R.N., Weiss, N. et al. Transgenic Expression and Functional Characterization of Human Platelet Derived Growth Factor BB (hPDGF-BB) in Tobacco (Nicotiana tabacum L.). Appl Biochem Biotechnol 171, 1390–1404 (2013). https://doi.org/10.1007/s12010-013-0413-x

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  • DOI: https://doi.org/10.1007/s12010-013-0413-x

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