International Journal of Industrial Entomology and Biomaterials

Korean Society of Sericultural Science (한국잠사학회)
권호 표지
DOI QR코드

DOI QR Code

Optimal Conditions for the Expression of Glycoprotein E2 of Classical Swine Fever Virus using Baculovirus in Insect Cells

  • Bae, Sung Min (Department of Agricultural Biology, College of Agriculture, Life and Environment Sciences, Chungbuk National University) ;
  • Lee, Seung Hee (Department of Agricultural Biology, College of Agriculture, Life and Environment Sciences, Chungbuk National University) ;
  • Kwak, Won Suk (Department of Agricultural Biology, College of Agriculture, Life and Environment Sciences, Chungbuk National University) ;
  • Ahn, Yong Oh (Department of Agricultural Biology, College of Agriculture, Life and Environment Sciences, Chungbuk National University) ;
  • Shin, Tae Young (Department of Agricultural Biology, College of Agriculture, Life and Environment Sciences, Chungbuk National University) ;
  • Woo, Soo Dong (Department of Agricultural Biology, College of Agriculture, Life and Environment Sciences, Chungbuk National University)
  • Received : 2014.12.26
  • Accepted : 2014.12.29
  • Published : 2014.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

The structural proteins of classical swine fever virus (CSFV) consist of nucleocapsid protein C and envelope glycoprotein $E^{rns}$ (E0), E1 and E2. Among them, E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. In this study, to determine the optimal expression conditions of glycoprotein E2 using baculovirus system, we investigated the influence of insect cells and media to the expression of recombinant E2. Recombinant virus containing glycoprotein E2 coding gene was constructed with bApGOZA DNA. Expression of the glycoprotein E2 was analyzed by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies. Expression of glycoprotein E2 in Sf21 cells was first observed after 3 days and reached a maximum on the 5th day after infection. Furthermore, the highest levels of glycoprotein E2 expression were observed at multiplicity of infection (MOI) of 5. When three different insect cell lines (Sf21, High-Five and Se301) were tested, High-Five cells showed the highest production. In addition, four different serum-free and serum-supplemented media, respectively, were tested for the expression of glycoprotein E2 and the budded virus (BV) titers. As a result, serum-supplemented medium provided the best conditions for protein production and the BV yield.

Keywords

References

  1. Ayres MD, Howard SC, Kuzio J, Lopez-Ferber M, Possee RD (1994) The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. Virology 202(2), 586-605. https://doi.org/10.1006/viro.1994.1380
  2. Bedard C, Kamen A, Tom R, Massie B (1994) Maximization of recombinant protein yield in the insect cell/baculovirus system by one-time addition of nutrients to high-density batch cultures. Cytotechnology, 15, 129-138. https://doi.org/10.1007/BF00762387
  3. Davis TR, Wickham TJ, McKenna KA, Granados RR, Shuler ML, Wood HA (1993) Comparative recombinant protein production of eight insect cell lines. In Vitro Cell. Dev. Biol. 29A, 388-390.
  4. Francki RIB, Fauquet CM, Knudson DL, Brown F (ed.) (1991) Flaviviridae, In Fifth report of the International Committee on Taxonomy of Viruses. p. 223-233, Springer Verlag, Vienna.
  5. Hink WF, Thomsen DR, Davidson DJ, Meyer AL, Castellino FJ (1991) Expression of three recombinant proteins using baculovirus vectors in 23 insect cell lines. Biotechnol. Prog.7(1), 9-14. https://doi.org/10.1021/bp00007a002
  6. Hulst MM, Westra DF, Wensvoort G, Moormann RJM (1993) Glycoprotein E1 of hog cholera virus expressed in insect cells protects swine from hog cholera. J. Virol. 67, 5435-5442.
  7. Ikonomou L, Schneider YJ, Agathos SN (2003) Insect cell culture for industrial production of recombinant proteins. Appl. Microbiol. Biotecnol. 62, 1-20. https://doi.org/10.1007/s00253-003-1223-9
  8. Je YH, Chang JH, Roh JY, Jin BR (2001) Generation of Baculovirus Expression Vector Using Defective Autographa californica Nuclear Polyhedrosis Virus Genome Maintained in Escherichia coli for Occ+ Virus Production. Int. J. Entomol. 2, 155-160.
  9. Jesionowski GA, Ataai MM (1997) An Efficient Medium for High Protein Production in the Insect Cell/Baculovirus Expression System, Biotechnol. Prog. 13, 355-360. https://doi.org/10.1021/bp970054l
  10. Kim JS, Choi JY, Roh JY, Lee HY, Jang SS, Je YH (2007) Production of recombinant polyhedral containing cry1Ac fusion protein in insect cell lines, J. Microbiol. Biotechnol. 17(5), 739-744.
  11. King LA, Possee RD (1992) The Baculovirus Expression System.A Laboratory Guide. Chapman & Hall, London.
  12. Lazarte JE, Tosi P, Nicolau C (1992) Optimization of the production of full-length rCD4 in baculovirus-infected Sf9 cells, Biotechnol. Bioeng. 40, 214-217. https://doi.org/10.1002/bit.260400204
  13. Licari P, Bailey JE (1991) Factors influencing recombinant protein yields in an insect cell-baculovirus expression system: multiplicity of infection and intracellular protein degradation, Biotechnol. Bioeng. 37, 238-246. https://doi.org/10.1002/bit.260370306
  14. Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4, Nature 227, 680-685. https://doi.org/10.1038/227680a0
  15. Lindsay DA, Betenbaugh MJ (1992) Quantification of cell culture factors affecting recombinant protein yields in baculovirus infected insect cells, Biotechnol. Bioeng. 39, 614-618. https://doi.org/10.1002/bit.260390605
  16. Matindoost L, Hu H, Chan LC, Nielsen LK, Reid S (2014) The effect of cell line, phylogenetics and medium on baculovirus budded virus yield and quality. Archives of virology, 159(1), 91-102. https://doi.org/10.1007/s00705-013-1789-1
  17. McIntosh AH, Grasela JJ (1994) Expression of $\beta$-galactosidase and luciferase in insect cell lines infected with a recombinant AcMNPV, In Vitro Cell. Dev. Biol. 30A, 275-278.
  18. Nishikawa N, Yamaji H, Fukuda H (2003) Recombinant protein production by the baculovirus-insect cell system in basal media without serum supplementation, Cytotechnology, 43, 3-10. https://doi.org/10.1023/B:CYTO.0000039894.27256.0f
  19. O'Reilly DR, Miller LK, Luckow VA (1992) Baculovirus Expression Vectors - A Laboratory Manual, W. H. Freeman and Company, New York.
  20. Paton DJ, McGoldrick A, Greiser-Wilke I, Parchariyanon S, Song JY, Liou PP, Stadejek T, Lowings JP, Bjorklund H, Belak S (2000) Genetic typing of classical swine fever virus, Vet. Microbiol. 73, 137-157. https://doi.org/10.1016/S0378-1135(00)00141-3
  21. Radford K, Cavegn C, Bertrand M, Bernard A, Reid S, Greenfield P (1997) The indirect effects of multiplicity of infection on baculovirus expressed proteins in insect cells: secreted and nonsecreted products, Cytotechnology, 24, 73-81. https://doi.org/10.1023/A:1007962903435
  22. Reuveny S, Kim YJ, Kemp CW, Shiloach J (1993) Effects of temperature and oxygen on cell growth and recombinant protein production in insect cell cultures, Appl. Microbiol. Biotechnol. 38, 619-623.
  23. Reuveny S, Kim YJ, Kemp CW, Shiloach J (1993) Production of recombinant proteins in high-density insect cell cultures, Biotechnol. Bioeng. 42, 235-239. https://doi.org/10.1002/bit.260420211
  24. Terpstra C (1991) Special review series: Hog cholera: an update of present knowledge, Br Vet J. 147, 397-406. https://doi.org/10.1016/0007-1935(91)90081-W
  25. Thiel HJ, Stark R, Weiland E, Rumenapf T, Meyers G (1991) Hog cholera virus: Molecular composition of virions from a pestivirus, J. Virol. 65, 4705-4712.
  26. van Rijn PA, van Gennip HGP, Moormann RJM (1999) An experimental marker vaccine and accompanying serological diagnostic test both based on envelope glycoprotein E2 of classical swine fever virus (CSFV), Vaccine17, 433-440. https://doi.org/10.1016/S0264-410X(98)00215-1
  27. van Zijl M, Wensvoort G, De Kluyver E, Hulst M, Van der Gulden H, Gielkens A, Gielkens A,Berns A, Moormann R (1991) Live attenuated pseudorabies virus expressing envelope glycoprotein E1 of hog cholera virus protects swine against both pseudorabies and hog cholera. J. Virol. 65(5), 2761-2765.
  28. Vaughn JL, Goodwin RH, Thompkins GJ, McCawley P (1977) The establishment of two insect cell lines from the insect S. frugiperda (Lepidoptera: Noctuidae), In Vitro. 13(4), 213-217. https://doi.org/10.1007/BF02615077
  29. Vlak J, Keus R (1990) Baculovirus expression vector system for production of viral vaccines, p. 91-128, In A. Mizrahi (ed.), Viral vaccines, Wiley-Liss, New York.
  30. Wickham TJ, Davis T, Granados RR, Hammer DA, Shuler ML, Wood HA (1991) Baculovirus defective interfering particles are responsible for variations in recombinant protein production as a function of multiplicity of infection, Biotechnol. Lett. 13, 483-488. https://doi.org/10.1007/BF01049204
  31. Wickham TJ, Davis T, Granados RR, Shuler ML, Wood HA (1992) Screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system, Biotechnol. Prog. 8(5), 391-396. https://doi.org/10.1021/bp00017a003
  32. Wong TK, Peter CH, Greenfield PF, Reid S, Nielsen LK (1996) Low multiplicity infection of insect cells with a recombinant baculovirus: the cell yield concept, Biotechnol. Bioeng. 49, 659-666.
  33. Wu J, King G, Daugulis AJ, Faulkner P, Bone DH, Goosen MFA (1989) Engineering aspects of insect cell suspension culture: a review. Appl. Microbiol. Biotecnol. 32(3), 249-255.