Skip to main content

Susceptibility to AcMNPV and expression of recombinant proteins by a novel cell clone derived from a Trichoplusia ni QAU-BTI-Tn9-4s cell line

Virologica Sinica volume 26, pages 297–305 (2011)Cite this article

Abstract

It is well known that Tn5B1-4 (commercially known as the High Five) cell line is highly susceptible to baculovirus and provides superior production of recombinant proteins when compared to other insect cell lines. But the characteristics of the cell line do not always remain stable and may change upon continuous passage. Recently an alphanodavirus, named Tn5 Cell Line Virus (or TNCL Virus), was identified in High Five cells in particular. Therefore, we established a new cell line, QB-Tn9-4s, from Trichoplusia ni, which was determined to be free of TNCL virus by RT-PCR analysis. In this paper, we describe the development of a novel cell clone, QB-CL-B, from a low passage QB-Tn9-4s cell line and report its susceptibility to AcMNPV, and the level of recombinant protein production. This cell clone was similar to its parental cells QB-Tn9-4s and Tn5B1-4 cells in morphology and growth rate; although it also showed approximately the same responses to AcMNPV infection and production of occlusion bodies, there were higher levels of recombinant protein production in comparison to QB-Tn9-4s (parental cells) and High5 cells.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

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

References

  1. Clemm D L. 1992. Scale-up of protein production in a stirred bioreactor. In: Baculovirus Expression Vectors, a Laboratory Manual. New York: W.H. Freeman and Company. p241–248.

    Google Scholar 

  2. Davis T R, Trotter K M, Granados R R, et al. 1992. Baculovirus expression of alkaline phosphatase as a reporter gene for evaluation of production, glycosylation and secretion. Biotechnology, 10: 1148–1150.

    Article  PubMed  CAS  Google Scholar 

  3. Davis T R, Wickham T J, Mckenna K A, et al. 1993. Comparative recombinant protein production of eight insect cell lines. In Vitro Cell Dev Biol-Anim, 29A: 388–390.

    Article  PubMed  CAS  Google Scholar 

  4. Donaldson M S, Shuler M L. 1998. Effects of long-term passaging of BTI-Tn-5B1-4 insect cells on growth and recombinant protein production. Biotechnol Progr, 14: 543–547.

    Article  CAS  Google Scholar 

  5. Granados R R, Li G, Blissard G W. 2007. Insect Cell Culture and Biotechnology. Virol Sin, 22(2): 83–93.

    Article  CAS  Google Scholar 

  6. Granados R R, Li G, Derksen A C G, et al. 1994. A new insect cell line from Trichoplusia ni (BTI-Tn-5B1-4) susceptible to Trichoplusia ni singly enveloped nuclear polyhedrosis virus. J Invertebr Pathol, 64: 260–266.

    Article  Google Scholar 

  7. Hayflick L. 1973. Theory of population increase by subcultivation.In: Tissue Culture Methods and Application. New York: Academic Press. p222–223.

    Google Scholar 

  8. Jarvis D L. 1997. Baculovirus expression vectors.In: The Baculoviruses. New York: Plenum Press. p389–431.

    Google Scholar 

  9. Li G, Hashimoto Y, Granados R R. 2003. Growth characteristics and expression of recombinant protein by new cell clones derived from Trichoplusia ni (BTI-Tn5B1-4) High Five cells. Bioprocessing, 2: 35–38.

    Google Scholar 

  10. Li T C, Scotti P D, Miyamura T, et al. 2007. Latent Infection of a New Alphanodavirus in an Insect Cell Line. J Virol, 81: 10890–10896.

    Article  PubMed  CAS  Google Scholar 

  11. Mckenna K A, Hong H, Vannunen E, et al. 1998. Establishment of new Trichoplusia ni cell lines in serum-free medium for baculovirus and recombinant protein production. J Invertebr Pathol, 71: 82–90.

    Article  CAS  Google Scholar 

  12. Meng M, Li T, Li C, et al. 2008. A suspended cell line from Trichoplusia ni (Lepidoptera): characterization and expression of recombinant proteins. Insect Sci, 15: 423–428.

    Article  CAS  Google Scholar 

  13. Wang P, Granados R R, Shuler M L. 1992. Studies on serum-free culture of insect cells for virus propagation and recombinant protein production. J Invertebr Pathol, 59: 46–53.

    Article  CAS  Google Scholar 

  14. Wang P, Toung R, Granados R R. 1999. The establishment of new cell lines from Pseudaletia unipuncta with differential responses to baculovirus infection. In Vitro Cell Dev Biol-Anim, 35: 333–338.

    Article  PubMed  CAS  Google Scholar 

  15. Wickham T J, Davis T, Granados R R, et al. 1992. Screening of insect cell lines for the production of recombinant and infectious virus in the baculovirus expression system. Biotechnol Progr, 8: 391–396.

    Article  CAS  Google Scholar 

  16. Wong K T K, Peter C H, Greenfield P F, et al. 1996. Low multiplicity infection of insect cells with a recombinant baculovirus: the cell yield concept. Biotechnol Bioeng, 49: 659–666.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. A Center for Advanced Invertebrate Cell Engineering Collaborated Between Boyce Thompson Institute and Qingdao Agricultural University, Qingdao Agricultural University (QAU), Qingdao, 266109, China

    Ming Shan, Lei Jiang, Ming Ma & Guo-xun Li

  2. Boyce Thompson Institute (BTI) at Cornell University, Ithaca, New York, 14853, USA

    Shi-ying Zhang

Authors
  1. Ming Shan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shi-ying Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lei Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ming Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Guo-xun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guo-xun Li.

Additional information

Foundation items: This research was supported in part by the Chinese National Basic Research Program (973) 2009CB118900, Chinese National Science Foundation Project 30771451, and Boyce Thompson Institute Project BTI-QAU 1-23-2007.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Shan, M., Zhang, Sy., Jiang, L. et al. Susceptibility to AcMNPV and expression of recombinant proteins by a novel cell clone derived from a Trichoplusia ni QAU-BTI-Tn9-4s cell line. Virol. Sin. 26, 297–305 (2011). https://doi.org/10.1007/s12250-011-3201-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12250-011-3201-1

Key words

  • Cell line
  • Insect virus
  • Recombinant protein expression
  • RT-PCR
  • TNCL virus