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Expression of hepatitis B small surface antigen in Santalum album embryogenic cell suspension cultures

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Biologia Plantarum

Abstract

Embryogenic cell suspension cultures of Santalum album were transformed with Agrobacterium tumefaciens harboring pD35SHER plant expression vector having hepatitis B small surface antigen (HBsAg) with a C-terminal ER retention signal. The transformed colonies were selected on culture medium supplemented with kanamycin and subsequently the transgenic nature of these colonies was confirmed by PCR analysis. The expression of HBsAg was confirmed by RT-PCR analysis and Western blot analysis and the expression was quantified using monoclonal antibody-based ELISA. Cell suspension cultures were initiated from the colony with expression of 11.09 μg(HBsAg) g−1(f.m.). To further increase the expression of HBsAg, transgenic S. album suspensions were cultured on media with various medium additives and cells growing in medium with 30 mM trehalose showed the expression of 19.95 μg(HBsAg) g−1(f.m.).

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Abbreviations

ELISA:

enzyme linked immunosorbent assay

HBsAg:

hepatitis B surface antigen

HER:

hepatitis B small surface antigen coding ‘s’ gene tagged with a C-terminal ER retention signal

RT-PCR:

reverse transcriptase-polymerase chain reaction

References

  • Čeřovská, N., Hoffmeisterová, H., Moravec, T., Plchová, H., Folwarczna, J., Hadámková, R.: Optimum storage conditions for product of transiently expressed epitopes of Human papillomavirus. — Biol. Plant. 52: 184–186, 2008.

    Article  Google Scholar 

  • Ganapathi, T.R., Sunil Kumar, G.B., Srinivas, L., Revathi, C.J., Bapat, V.A.: Analysis of the limitations of hepatitis B surface antigen expression in soybean cell suspension cultures. — Plant Cell Rep. 26: 1575–1584, 2007.

    Article  CAS  PubMed  Google Scholar 

  • Harrington, H.M., Berry, S.L., Henke, R.R.: Amino acid transport into cultured tobacco cells II. Effect of calcium. — Plant Physiol. 67: 379–384, 1981.

    Article  CAS  PubMed  Google Scholar 

  • Hellwig, S., Drossard, J., Twyman, R.M., Fischer, R.: Plant cell cultures for the production of recombinant proteins. — Nat. Biotechnol. 22: 1415–1422, 2004.

    Article  CAS  PubMed  Google Scholar 

  • Huang, J., Sutliff, T.D., Wu, L., Nandi, S., Benge, K., Terashima, M., Ralston, A.H., Drohan, W., Huang, N., Rodriguez, R.L.: Expression and purification of functional human alpha-1-antitrypsin from cultured plant cells. — Biotechnol. Progr. 17: 126–133, 2001.

    Article  CAS  Google Scholar 

  • Jaganathan, K.S., Singh, P., Prabakaran, D., Mishra, V., Vyas, S.P.: Development of a single-dose stabilized poly(D,Llactic-co-glycolic acid) microspheres-based vaccine against hepatitis B. — J. Pharm. Pharmacol. 56: 1243–1250, 2004.

    Article  CAS  PubMed  Google Scholar 

  • Kaushik, J.K., Bhat, R.: Why is trehalose an exceptional protein stabilizer? An analysis of the thermal stability of proteins in the presence of the compatible osmolyte trehalose. — J. biol. Chem. 278: 26458–26465, 2003.

    Article  CAS  PubMed  Google Scholar 

  • Kwon, T.H., Kim, Y.S., Lee, J.H., Yang, M.S.: Production and secretion of biologically active human granulocyte-macrophage colony stimulating factor in transgenic tomato suspension cultures. — Biotechnol. Lett. 25: 1571–1574, 2003.

    Article  CAS  PubMed  Google Scholar 

  • Lee, J.H., Kim, N.S., Kwon, T.H., Yang, M.S.: Effects of osmotic pressure on production of recombinant human granulocyte-macrophage colony stimulating factor in plant cell suspension culture. — Enzyme Microb. Technol. 30: 768–773, 2002.

    Article  CAS  Google Scholar 

  • Lhernould, S., Labrousse, P., Lejolly, D., Léonard, R., Carlué, M., Costa, G.: In vivo activity of recombinant human Lewis fucosyltransferase III in leaves of Nicotiana tabacum L. — Biol. Plant. 52: 267–274, 2008.

    Article  CAS  Google Scholar 

  • Maa, Y.F., Ameri, M., Shu, C., Zuleger, C.L., Che. J., Osorio, J.E., Payne, L.G., Chen, D.: Hepatitis-B surface antigen (HBsAg) powder formulation: process and stability assessment. — Curr. Drug Delivery 4: 57–67, 2007.

    Article  CAS  Google Scholar 

  • Negrouk, V., Eisner, G., Lee, H., Han, K., Taylor, D., Wong, H.C.: Highly efficient transient expression of functional recombinant antibodies in lettuce. — Plant Sci. 169: 433–438, 2005.

    Article  CAS  Google Scholar 

  • Pal, S., Das, S., Dey, S.: Peroxidase and arabinogalactan protein as by-products during somatic embryo cultivation in air-lift bioreactor. — Process Biochem. 38: 1471–1477, 2003.

    Article  CAS  Google Scholar 

  • Richter, L.J., Thanavala, Y., Arntzen, C.J., Mason, H.S.: Production of hepatitis B surface antigen in transgenic plants for oral immunization. — Nat. Biotechnol. 18: 1167–1171, 2000.

    Article  CAS  PubMed  Google Scholar 

  • Shekhawat, U.K.S., Ganapathi, T.R., Sunil Kumar, G.B., Srinivas, L.: Sucrose-inducible expression of hepatitis B surface antigen using potato granule-bound starch synthase promoter. — Plant Biotechnol. Rep. 1: 199–206, 2007.

    Article  Google Scholar 

  • Shekhawat, U.K.S., Ganapathi, T.R., Srinivas, L., Bapat, V.A., Rathore, T.S.: Agrobacterium-mediated genetic transformation of embryogenic cell suspension cultures of Santalum album L. — Plant Cell Tissue Organ Cult. 92: 261–271, 2008.

    Article  CAS  Google Scholar 

  • Sijmons, P.C., Dekker, B.M., Schrammeijer, B., Verwoerd, T.C., Van den Elzen, P.J., Hoekema, A.: Production of correctly processed human serum albumin in transgenic plants. — Bio/Technology 8: 217–221, 1990.

    Article  CAS  PubMed  Google Scholar 

  • Smith, M.L., Mason, H.S., Shuler, M.L.: Hepatitis B surface antigen (HBsAg) expression in plant cell culture: Kinetics of antigen accumulation in batch culture and its intracellular form. — Biotechnol. Bioeng. 80: 812–822, 2002.

    Article  CAS  PubMed  Google Scholar 

  • Soderquist, R.G., Lee, J.M.: Enhanced production of recombinant proteins from plant cells by the application of osmotic stress and protein stabilization. — Plant Cell Rep. 24: 127–132, 2005.

    Article  CAS  PubMed  Google Scholar 

  • Streatfield, S.J.: Oral hepatitis B vaccine candidates produced and delivered in plant material. — Immunol. Cell Biol. 83: 257–262, 2005.

    Article  CAS  PubMed  Google Scholar 

  • Sunil Kumar, G.B., Ganapathi, T.R., Bapat, V.A.: Production of hepatitis B surface antigen in recombinant plant systems: an update. — Biotechnol. Prog. 23: 532–539, 2007.

    Article  Google Scholar 

  • Tanaka, R.D., Li, A.C., Fogelman, A.M., Edwards, P.A.: Inhibition of lysosomal protein degradation inhibits the basal degradation of 3-hydroxy-3-methylglutaryl coenzyme A reductase. — J. Lipid Res. 27: 261–273, 1986.

    CAS  PubMed  Google Scholar 

  • Tsoi, B.M., Doran, P.M.: Effect of medium properties and additives on antibody stability and accumulation in suspended plant cell cultures. — Biotechnol. appl. Biochem. 35: 171–180, 2002.

    Article  CAS  PubMed  Google Scholar 

  • Whal, M.F., An, G.H.A., Lee, J.M.: Effects of dimethyl sulfoxide on heavy chain monoclonal antibody production from plant cell culture. — Biotechnol. Lett. 17: 463–468, 1995.

    Article  Google Scholar 

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Acknowledgements

Authors thank Dr C.J. Revathi, Shantha Biotechnics Ltd, Hyderabad, for providing the ’s’ gene coding for HBsAg.

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Authors and Affiliations

  1. Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085, India

    U. K. S. Shekhawat, T. R. Ganapathi & L. Srinivas

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  1. U. K. S. Shekhawat
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  2. T. R. Ganapathi
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  3. L. Srinivas
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Correspondence to T. R. Ganapathi.

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Shekhawat, U.K.S., Ganapathi, T.R. & Srinivas, L. Expression of hepatitis B small surface antigen in Santalum album embryogenic cell suspension cultures. Biol Plant 54, 720–724 (2010). https://doi.org/10.1007/s10535-010-0128-6

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  • DOI: https://doi.org/10.1007/s10535-010-0128-6

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