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Transgenic Research

, Volume 24, Issue 5, pp 897–909 | Cite as

Tobacco seeds as efficient production platform for a biologically active anti-HBsAg monoclonal antibody

  • Abel Hernández-Velázquez
  • Alina López-Quesada
  • Yanaysi Ceballo-Cámara
  • Gleysin Cabrera-Herrera
  • Kenia Tiel-González
  • Liliana Mirabal-Ortega
  • Marlene Pérez-Martínez
  • Rosabel Pérez-Castillo
  • Yamilka Rosabal-Ayán
  • Osmani Ramos-González
  • Gil Enríquez-Obregón
  • Ann Depicker
  • Merardo Pujol-Ferrer
Original Paper

Abstract

The use of plants as heterologous hosts is one of the most promising technologies for manufacturing valuable recombinant proteins. Plant seeds, in particular, constitute ideal production platforms for long-term applications requiring a steady supply of starting material, as they combine the general advantages of plants as bioreactors with the possibility of biomass storage for long periods in a relatively small volume, thus allowing manufacturers to decouple upstream and downstream processing. In the present work we have used transgenic tobacco seeds to produce large amounts of a functionally active mouse monoclonal antibody against the Hepatitis B Virus surface antigen, fused to a KDEL endoplasmic reticulum retrieval motif, under control of regulatory sequences from common bean (Phaseolus vulgaris) seed storage proteins. The antibody accumulated to levels of 6.5 mg/g of seed in the T3 generation, and was purified by Protein A affinity chromatography combined with SEC-HPLC. N-glycan analysis indicated that, despite the KDEL signal, the seed-derived plantibody bore both high-mannose and complex-type sugars that indicate partial passage through the Golgi compartment, although its performance in the immunoaffinity purification of HBsAg was unaffected. An analysis discussing the industrial feasibility of replacing the currently used tobacco leaf-derived plantibody with this seed-derived variant is also presented.

Keywords

Protein expression Tobacco seeds Phaseolin promoter Monoclonal antibody Hepatitis B N-glycan 

Notes

Acknowledgments

We thank the Department of Plant Systems Biology of Ghent University Belgium for supplying the signals for expression in seeds. In addition, we would like to extend our appreciation to the staff of the experimental area at CIGB for their help in cultivating the tobacco plants used in this study and to the Monoclonal Antibody Production Department at CIGB for their help with the immunoaffinity purification of HBsAg. No potential conflicts of interest are declared in this work.

Supplementary material

11248_2015_9890_MOESM1_ESM.docx (403 kb)
Supplementary material 1 (DOCX 403 kb)

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Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Abel Hernández-Velázquez
    • 1
  • Alina López-Quesada
    • 1
  • Yanaysi Ceballo-Cámara
    • 1
  • Gleysin Cabrera-Herrera
    • 2
  • Kenia Tiel-González
    • 1
  • Liliana Mirabal-Ortega
    • 1
  • Marlene Pérez-Martínez
    • 1
  • Rosabel Pérez-Castillo
    • 1
  • Yamilka Rosabal-Ayán
    • 1
  • Osmani Ramos-González
    • 1
  • Gil Enríquez-Obregón
    • 1
  • Ann Depicker
    • 3
  • Merardo Pujol-Ferrer
    • 1
  1. 1.Plant Biotechnology DepartmentCenter for Genetic Engineering and Biotechnology (CIGB)HavanaCuba
  2. 2.Department of Carbohydrate ChemistryCenter for Genetic Engineering and Biotechnology (CIGB)HavanaCuba
  3. 3.Department of Plant Systems BiologyVIB, Plant-made Antibodies and ImmunogensGhentBelgium

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