Skip to main content

Advertisement

SpringerLink
Log in

Transient expression of human serum albumin (HSA) in tobacco leaves

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

A Correction to this article was published on 07 September 2020

This article has been updated

Abstract

Today, recombinant human proteins make up a considerable part of FDA-approved biotechnological drugs. The selection of proper expression platform for manufacturing recombinant protein is a vital factor in achieving the optimal yield and quality of a biopharmaceutical in a timely fashion. This experiment was aimed to compare the transient expression level of human serum albumin gene in different tobacco genotype. For this, the Agrobacterium tumefaciens strains LB4404 and GV3101 harboring pBI121-HSA binary vector were infiltered in leaves of three tobacco genotypes, including Nicotiana benthamiana and N. tabacum cv Xanthi and Samsun. The qRT-PCR, SDS-PAGE, western blotting and ELISA analysis were performed to evaluate the expression of HSA gene in transgenic plantlets. Our results illustrated that the expression level of rHSA in tobacco leaves was highly dependent on Agrobacterium strains, plant genotypes and harvesting time. The highest production of recombinant HSA protein was obtained in Samsun leaves infected with A. tumefaciens strain GV3101 after 3 days of infiltration.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Change history

  • 07 September 2020

    The article ‘Transient expression of human serum albumin (HSA) in tobacco leaves’ written by Behnam Sedaghati, Raheem Haddad, and Mojgan Bandehpour, was originally published online on 8th July 2020 with Open Access under a Creative Commons Attribution (CC BY) license 4.0. With the authors' decision to cancel Open Access the copyright of the article changed on 21st August 2020 to @Springer Nature B.V. 2020 with all rights reserved. The original article has been corrected.

References

  1. Barbosa S, Taboada P, Mosquera V (2014) Chapter 32—fibrillation and polymorphism of human serum albumin. In: Uversky VN, Lyubchenko YL (eds) Bio-nanoimaging. Academic Press, Boston, pp 345–362

    Google Scholar 

  2. Swiech K, Picanço-Castro V, Covas DT (2012) Human cells: new platform for recombinant therapeutic protein production. Protein Express Purif 84:147–153

    CAS  Google Scholar 

  3. Xu J, Towler M, Weathers PJ (2016) Platforms for plant-based protein production. In: Pavlov A, Bley T (eds) Bioprocessing of plant in vitro systems. Springer, Cham, pp 1–40

    Google Scholar 

  4. Ma L, Lukasik E, Gawehns F, Takken FL (2012) The use of agroinfiltration for transient expression of plant resistance and fungal effector proteins in Nicotiana benthamiana leaves. In: Ma L, Lukasik E, Gawehns F, Takken FLW (eds) Plant fungal pathogens. Springer, New York, pp 61–74

    Google Scholar 

  5. Sedaghati B, Haddad R, Bandehpour M (2019) Efficient plant regeneration and Agrobacterium-mediated transformation via somatic embryogenesis in purslane (Portulaca oleracea L.): an important medicinal plant. Plant Cell Tissue Org 136:231–245

    CAS  Google Scholar 

  6. Japelaghi RH, Haddad R, Garoosi G-A (2011) Rapid and efficient isolation of high quality nucleic acids from plant tissues rich in polyphenols and polysaccharides. Mol Biotechnol 49:129–137

    CAS  PubMed  Google Scholar 

  7. Snell EJ, Simpson H (1991) Applied statistics: handbook of GENSTAT analysis, vol 13. CRC Press, Boca Raton

    Google Scholar 

  8. Chen Q, Davis KR (2016) The potential of plants as a system for the development and production of human biologics. F1000Res 5

  9. Buyel JF (2019) Plant molecular farming—integration and exploitation of side streams to achieve sustainable biomanufacturing. Front Plant Sci 9:1893–1893

    PubMed  PubMed Central  Google Scholar 

  10. Lau OS, Sun SS (2009) Plant seeds as bioreactors for recombinant protein production. Biotechnol Adv 27:1015–1022

    CAS  PubMed  Google Scholar 

  11. Matoba N, Davis KR, Palmer KE (2011) Recombinant protein expression in nicotiana. In: Birchler JA (ed) Plant chromosome engineering: methods and protocols. Humana Press, Totowa, pp 199–219

    Google Scholar 

  12. Tremblay R, Wang D, Jevnikar AM, Ma S (2010) Tobacco, a highly efficient green bioreactor for production of therapeutic proteins. Biotechnol Adv 28:214–221

    CAS  PubMed  Google Scholar 

  13. Vojta L, Ljuma-Skupnjak L, Budimir A, Vukičević S, Fulgosi H (2015) Rapid transient expression of human granulocyte-macrophage colony-stimulating factor in two industrial cultivars of tobacco (Nicotiana tabacum L.) by agroinfiltration. Biotechnol Rep 7:81–86

    Google Scholar 

  14. Xu J, Okada S, Tan L, Goodrum KJ, Kopchick JJ, Kieliszewski MJ (2010) Human growth hormone expressed in tobacco cells as an arabinogalactan-protein fusion glycoprotein has a prolonged serum life. Transgenic Res 19:849–867

    CAS  PubMed  Google Scholar 

  15. Wirth S, Calamante G, Mentaberry A, Bussmann L, Lattanzi M, Barañao L, Bravo-Almonacid F (2004) Expression of active human epidermal growth factor (hEGF) in tobacco plants by integrative and non-integrative systems. Mol Breed 13:23–35

    CAS  Google Scholar 

  16. Valdés R, Gómez L, Padilla S, Brito J, Reyes B, Álvarez T, Mendoza O, Herrera O, Ferro W, Pujol M, Leal V, Linares M, Hevia Y, Garcı́a C, Milá L, Garcı́a O, Sánchez R, Acosta A, Geada D, Paez R, Luis Vega J, Borroto C (2003) Large-scale purification of an antibody directed against hepatitis B surface antigen from transgenic tobacco plants. Biochem Biophys Res Commun 308:94–100

    PubMed  Google Scholar 

  17. Nakashima K, Shibasaki-Kitakawa N, Miyamoto T, Kubo M, Yonemoto T, Shuler ML (2013) Production of human secreted alkaline phosphatase in suspension and immobilization cultures of tobacco NT1 cell. Biochem Eng J 77:177–182

    CAS  Google Scholar 

  18. Shah KH, Almaghrabi B, Bohlmann H (2013) Comparison of expression vectors for transient expression of recombinant proteins in plants. Plant Mol Biol Rep 31:1529–1538

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Krenek P, Samajova O, Luptovciak I, Doskocilova A, Komis G, Samaj J (2015) Transient plant transformation mediated by Agrobacterium tumefaciens: principles, methods and applications. Biotechnol Adv 33:1024–1042

    CAS  PubMed  Google Scholar 

  20. Yusibov V, Streatfield SJ, Kushnir N (2011) Clinical development of plant-produced recombinant pharmaceuticals: vaccines, antibodies and beyond. Hum Vaccines 7:313–321

    CAS  Google Scholar 

  21. Vaghchhipawala Z, Rojas CM, Senthil-Kumar M, Mysore KS (2011) Agroinoculation and Agroinfiltration: simple tools for complex gene function analyses. In: Pereira A (ed) Plant reverse genetics: methods and protocols. Humana Press, Totowa, pp 65–76

    Google Scholar 

  22. Wu H-Y, Liu K-H, Wang Y-C, Wu J-F, Chiu W-L, Chen C-Y, Wu S-H, Sheen J, Lai E-M (2014) AGROBEST: an efficient Agrobacterium-mediated transient expression method for versatile gene function analyses in Arabidopsis seedlings. Plant Methods 10:19

    PubMed  PubMed Central  Google Scholar 

  23. Bidarigh fard A, Dehghan Nayeri F, Habibi Anbuhi M (2019) Transient expression of etanercept therapeutic protein in tobacco (Nicotiana tabacum L.). Int J Biol Macromol 130:483–490

    CAS  PubMed  Google Scholar 

  24. Dickey A, Wang N, Cooper E, Tull L, Breedlove D, Mason H, Liu D, Wang KY (2017) Transient expression of Lumbrokinase (PI239) in tobacco (Nicotiana tabacum) using a geminivirus-based single replicon system dissolves fibrin and blood clots. Evid-Based Compl Altern 2017:1–9

    Google Scholar 

  25. Teh Y-HA, Kavanagh TA (2010) High-level expression of Camelid nanobodies in Nicotiana benthamiana. Transgenic Res 19:575–586

    CAS  PubMed  Google Scholar 

  26. Feng Z-G, Pang S-F, Guo D-J, Yang Y-T, Liu B, Wang J-W, Zheng K-Q, Lin Y (2014) Recombinant keratinocyte growth factor 1 in tobacco potentially promotes wound healing in diabetic rats. Biomed Res Int 2014:1–10

    Google Scholar 

  27. Bhaskar PB, Venkateshwaran M, Wu L, Ané J-M, Jiang J (2009) Agrobacterium-mediated transient gene expression and silencing: a rapid tool for functional gene assay in potato. PLoS ONE 4(e5812):1–8

    Google Scholar 

  28. Wu S-L, Yang X-B, Liu L-Q, Jiang T, Wu H, Su C, Qian Y-H, Jiao F (2015) Agrobacterium-mediated transient MaFT expression in mulberry (Morus alba L.) leaves. Biosci Biotechnol Biochem 79:1266–1271

    CAS  PubMed  Google Scholar 

  29. Kim MJ, Baek K, Park C-M (2009) Optimization of conditions for transient Agrobacterium-mediated gene expression assays in Arabidopsis. Plant Cell Rep 28:1159–1167

    CAS  PubMed  Google Scholar 

  30. Norkunas K, Harding R, Dale J, Dugdale B (2018) Improving agroinfiltration-based transient gene expression in Nicotiana benthamiana. Plant Methods 14:1–14

    Google Scholar 

  31. Cao DV, Pamplona RS, Kim J, Oh YK, Cho S-K, Ahn J, Yang S-W, Riu K-Z, Boo K-H (2017) Optimization of Agrobacterium-mediated transient expression of heterologous genes in spinach. Plant Biotechnol Rep 11:397–405

    Google Scholar 

  32. Vargas-Guevara C, Vargas-Segura C, Villalta-Villalobos J, Pereira LF, Gatica-Arias A (2018) A simple and efficient agroinfiltration method in coffee leaves (Coffea arabica L.): assessment of factors affecting transgene expression. 3 Biotech 8:471

    PubMed  PubMed Central  Google Scholar 

  33. Conley AJ, Zhu H, Le LC, Jevnikar AM, Lee BH, Brandle JE, Menassa R (2011) Recombinant protein production in a variety of Nicotiana hosts: a comparative analysis. Plant Biotechnol J 9:434–444

    CAS  PubMed  Google Scholar 

  34. Citovsky V, Kozlovsky SV, Lacroix B, Zaltsman A, Dafny-Yelin M, Vyas S, Tovkach A, Tzfira T (2007) Biological systems of the host cell involved in Agrobacterium infection. Cell Microbiol 9:9–20

    CAS  PubMed  Google Scholar 

  35. Gohlke J, Deeken R (2014) Plant responses to Agrobacterium tumefaciens and crown gall development. Front Plant Sci 5:155–155

    PubMed  PubMed Central  Google Scholar 

  36. Karami O, Esna-Ashari M, Kurdistani GK, Aghavaisi B (2009) Agrobacterium-mediated genetic transformation of plants: the role of host. Biol Plant 53:201–212

    CAS  Google Scholar 

  37. Yasmin A, Debener T (2010) Transient gene expression in rose petals via Agrobacterium infiltration. Plant Cell Tissue Org 102:245–250

    CAS  Google Scholar 

  38. Zhao Q, Du Y, Wang H, Rogers HJ, Yu C, Liu W, Zhao M, Xie F (2019) 5-Azacytidine promotes shoot regeneration during Agrobacterium-mediated soybean transformation. Plant Physiol Biochnol 141:40–50

    CAS  Google Scholar 

  39. Mo R, Huang Y, Yang S, Zhang Q, Luo Z (2015) Development of Agrobacterium-mediated transient transformation in persimmon (Diospyros kaki Thunb.). Sci Hortic 192:29–37

    CAS  Google Scholar 

  40. Santos-Rosa M, Poutaraud A, Merdinoglu D, Mestre P (2008) Development of a transient expression system in grapevine via agro-infiltration. Plant Cell Rep 27:1053–1063

    CAS  PubMed  Google Scholar 

  41. Zottini M, Barizza E, Costa A, Formentin E, Ruberti C, Carimi F, Schiavo FL (2008) Agroinfiltration of grapevine leaves for fast transient assays of gene expression and for long-term production of stable transformed cells. Plant Cell Rep 27:845–853

    CAS  PubMed  Google Scholar 

  42. Faizal A, Geelen D (2012) Agroinfiltration of intact leaves as a method for the transient and stable transformation of saponin producing Maesa lanceolate. Plant Cell Rep 31:1517–1526

    CAS  PubMed  Google Scholar 

  43. Sheludko Y, Sindarovska Y, Gerasymenko I, Bannikova M, Kuchuk N (2007) Comparison of several Nicotiana species as hosts for high-scale Agrobacterium-mediated transient expression. Biotechnol Bioeng 96:608–614

    CAS  PubMed  Google Scholar 

  44. Chicas A, Macino G (2001) Characteristics of post-transcriptional gene silencing. EMBO Rep 2:992–996

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Laguia-Becher M, Zaldúa Z, Xu W, Marconi PL, Velander W, Alvarez MA (2019) Co-expressing Turnip Crinkle Virus-coat protein with the serine protease α-thrombin precursor (pFIIa) in Nicotiana benthamiana Domin. Vitro Cell Dev Biol 55:88–98

    CAS  Google Scholar 

  46. Sun QY, Ding LW, Lomonossoff GP, Sun YB, Luo M, Li CQ, Jiang L, Xu ZF (2011) Improved expression and purification of recombinant human serum albumin from transgenic tobacco suspension culture. J Biotechnol 155:164–172

    CAS  PubMed  Google Scholar 

  47. Fernández-San Millán A, Mingo-Castel A, Miller M, Daniell H (2003) A chloroplast transgenic approach to hyper-express and purify Human Serum Albumin, a protein highly susceptible to proteolytic degradation. Plant Biotechnol J 1:71–79

    PubMed  Google Scholar 

  48. Farran I, Sánchez-Serrano JJ, Medina JF, Prieto J, Mingo-Castel AM (2002) Targeted expression of human serum albumin to potato tubers. Transgenic Res 11:337–346

    CAS  PubMed  Google Scholar 

  49. Chen Z, He Y, Shi B, Yang D (2013) Human serum albumin from recombinant DNA technology: challenges and strategies. Biochim Biophys Acta 1830:5515–5525

    CAS  PubMed  Google Scholar 

  50. Sijmons PC, Dekker BM, Schrammeijer B, Verwoerd TC, Van Den Elzen PJ, Hoekema A (1990) Production of correctly processed human serum albumin in transgenic plants. Bio/Technology 8:217–221

    CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by Cellular and Molecular Biology Research Center at Shahid Beheshti University of Medical Sciences (Grant No. 22777).

Author information

Authors and Affiliations

  1. Department of Biotechnology, Faculty of Agriculture and Natural Resources, Imam Khomeini International University, Qazvin, Iran

    Behnam Sedaghati & Raheem Haddad

  2. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Behnam Sedaghati & Mojgan Bandehpour

Authors
  1. Behnam Sedaghati
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Raheem Haddad
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mojgan Bandehpour
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

BS: conceptualization, methodology, investigation, writing—original draft. RH: supervision, writing—review and editing. MB: investigation, writing—review and editing.

Corresponding author

Correspondence to Raheem Haddad.

Ethics declarations

Conflict of interest

The authors of the paper declare that there are no conflicts of interest.

Research involving human and animal rights

This article contains no studies with human or animal subjects performed by any of the authors.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

The original version of this article was revised due to a retrospective Open Access cancellation.

The article ‘Transient expression of human serum albumin (HSA) in tobacco leaves’ written by Behnam Sedaghati, Raheem Haddad and Mojgan Bandehpour was originally published online on 8th July 2020 with Open Access under a Creative Commons Attribution (CC BY) license 4.0. Withe the authors’ decision to cancel Open Access the copyright of the article changed on 21st August 2020 to @Springer Nature B.V. 2020 with all rights reserved. The original article has been corrected.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sedaghati, B., Haddad, R. & Bandehpour, M. Transient expression of human serum albumin (HSA) in tobacco leaves. Mol Biol Rep 47, 7169–7177 (2020). https://doi.org/10.1007/s11033-020-05640-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-020-05640-y

Keywords

Search

Navigation