The M4 insulator, the TM2 matrix attachment region, and the double copy of the heavy chain gene contribute to the enhanced accumulation of the PHB-01 antibody in tobacco plants

  • Yoslaine RuizEmail author
  • Pedro Luis Ramos
  • Jeny Soto
  • Meilyn Rodríguez
  • Natacha Carlos
  • Aneisi Reyes
  • Danay Callard
  • Yadira Sánchez
  • Merardo Pujol
  • Alejandro FuentesEmail author
Original Paper


The expression of recombinant proteins in plants is a valuable alternative to bioreactors using mammalian cell systems. Ease of scaling, and their inability to host human pathogens, enhance the use of plants to generate complex therapeutic products such as monoclonal antibodies. However, stably transformed plants expressing antibodies normally have a poor accumulation of these proteins that probably arise from the negative positional effects of their flanking chromatin. The induction of boundaries between the transgenes and the surrounding DNA using matrix attachment regions (MAR) and insulator elements may minimize these effects. With the PHB-01 antibody as a model, we demonstrated that the insertion of DNA elements, the TM2 (MAR) and M4 insulator, flanking the transcriptional cassettes that encode the light and heavy chains of the PHB-01 antibody, increased the protein accumulation that remained stable in the first plant progeny. The M4 insulator had a stronger effect than the TM2, with over a twofold increase compared to the standard construction. This effect was probably associated with an enhancer-promoter interference. Moreover, transgenic plants harboring two transcriptional units encoding for the PHB-01 heavy chain combined with both TM2 and M4 elements enhanced the accumulation of the antibody. In summary, the M4 combined with a double transcriptional unit of the heavy chain may be a suitable strategy for potentiating PHB-01 production in tobacco plants.


Antibody Plant expression Insulator Matrix attachment region Tobacco 



Matrix attachment region


Light chain


Heavy chain


Cauliflower mosaic virus



The authors thank Dra. Miriam Ribas Hermelo for critical review of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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© Springer Nature Switzerland AG 2020

Authors and Affiliations

  1. 1.Plant Biotechnology DepartmentCenter for Genetic Engineering and Biotechnology, CubaHavanaCuba
  2. 2.Department of Phytopathology and Plant BiochemistryInstituto BiologicoSão PauloBrazil
  3. 3.Comparative Pathology DepartmentUniversity of MiamiMiamiUSA

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