Preincubation with a low concentration of methyl-β-cyclodextrin enhances baculovirus expression system productivity

  • Jinshan Huang
  • Na Liu
  • Xingjia Shen
  • Bifang HaoEmail author
Original Research Paper



To enhance the productivity of foreign protein in culture cells using baculovirus expression system.


A low concentration of MβCD, with the optimal application concentration of 0.25 mM and the appropriate preincubation time range from 10 to 120 min, can efficiently enhance expression levels in both the AcMNPV and BmNPV expression systems.


Preincubation with a low concentration MβCD enhance baculovirus infection and foreign protein expression productivity.


AcMNPV BmNPV Expression Methyl-β-cyclodextrin 



This study was funded by the National Science Foundation of China (Grant No. 31670152); and the Scientific Research Foundation for Returned Overseas Chinese Scholars, State Education Ministry (Grant No. 2015).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. Bajimaya S, Hayashi T, Frankl T, Bryk P, Ward B, Takimoto T (2017) Cholesterol reducing agents inhibit assembly of type I parainfluenza viruses. Virology 501:127–135. CrossRefGoogle Scholar
  2. Boublik Y, Di Bonito P, Jones IM (1995) Eukaryotic virus display: engineering the major surface glycoprotein of the Autographa californica nuclear polyhedrosis virus (AcNPV) for the presentation of foreign proteins on the virus surface. Biotechnology (N Y) 13:1079–1084CrossRefGoogle Scholar
  3. Chaabihi H, Ogliastro MH, Martin M, Giraud C, Devauchelle G, Cerutti M (1993) Competition between baculovirus polyhedrin and p10 gene expression during infection of insect cells. J Virol 67:2664–2671Google Scholar
  4. Coutinho A, Silva L, Fedorov A, Prieto M (2004) Cholesterol and ergosterol influence nystatin surface aggregation: relation to pore formation. Biophys J 87:3264–3276. CrossRefGoogle Scholar
  5. Dong S, Wang M, Qiu Z, Deng F, Vlak JM, Hu Z, Wang H (2010) Autographa californica multicapsid nucleopolyhedrovirus efficiently infects Sf9 cells and transduces mammalian cells via direct fusion with the plasma membrane at low pH. J Virol 84:5351–5359. CrossRefGoogle Scholar
  6. Gomez-Sebastian S, Lopez-Vidal J, Escribano JM (2014) Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette. PLoS ONE 9:e96562. CrossRefGoogle Scholar
  7. Grimmer S, van Deurs B, Sandvig K (2002) Membrane ruffling and macropinocytosis in A431 cells require cholesterol. J Cell Sci 115:2953–2962Google Scholar
  8. Hitchman RB, Locanto E, Possee RD, King LA (2011) Optimizing the baculovirus expression vector system Methods 55:52–57. Google Scholar
  9. Huang J, Hao B, Sun X, Deng F, Wang H, Hu Z (2007) Construction of the Bac-to-Bac System of Bombyx mori Nucleopolyhedrovirus. Virol Sin 22:218–225CrossRefGoogle Scholar
  10. Huang J, Hao B, Cheng C, Liang F, Shen X, Cheng X (2014) Entry of Bombyx mori nucleopolyhedrovirus into BmN cells by cholesterol-dependent macropinocytic endocytosis. Biochem Biophys Res Commun 453:166–171. CrossRefGoogle Scholar
  11. Ilangumaran S, Hoessli DC (1998) Effects of cholesterol depletion by cyclodextrin on the sphingolipid microdomains of the plasma membrane. Biochem J 335(Pt 2):433–440CrossRefGoogle Scholar
  12. Je YH, Chang JH, Choi JY, Roh JY, Jin BR, O’Reilly DR, Kang SK (2001) A defective viral genome maintained in Escherichia coli for the generation of baculovirus expression vectors. Biotech Lett 23:575–582CrossRefGoogle Scholar
  13. Kataoka C et al (2012) Baculovirus GP64-mediated entry into mammalian cells. J Virol 86:2610–2620. CrossRefGoogle Scholar
  14. Katou Y, Yamada H, Ikeda M, Kobayashi M (2010) A single amino acid substitution modulates low-pH-triggered membrane fusion of GP64 protein in Autographa californica and Bombyx mori nucleopolyhedroviruses. Virology 404:204–214. CrossRefGoogle Scholar
  15. Klein U, Gimpl G, Fahrenholz F (1995) Alteration of the myometrial plasma membrane cholesterol content with beta-cyclodextrin modulates the binding affinity of the oxytocin receptor. Biochemistry 34:13784–13793CrossRefGoogle Scholar
  16. Kwik J, Boyle S, Fooksman D, Margolis L, Sheetz MP, Edidin M (2003) Membrane cholesterol, lateral mobility, and the phosphatidylinositol 4,5-bisphosphate-dependent organization of cell actin. Proc Natl Acad Sci USA 100:13964–13969. CrossRefGoogle Scholar
  17. Lee KS, Je YH, Woo SD, Sohn HD, Jin BR (2006) Production of a cellulase in silkworm larvae using a recombinant Bombyx mori nucleopolyhedrovirus lacking the virus-encoded chitinase and cathepsin genes. Biotechnol Lett 28:645–650. CrossRefGoogle Scholar
  18. Luckow VA, Lee SC, Barry GF, Olins PO (1993) Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli. J Virol 67:4566–4579Google Scholar
  19. Luz-Madrigal A, Asanov A, Camacho-Zarco AR, Sampieri A, Vaca L (2013) A cholesterol recognition amino acid consensus domain in GP64 fusion protein facilitates anchoring of baculovirus to mammalian cells. J Virol 87:11894–11907. CrossRefGoogle Scholar
  20. Maeda S et al (1985) Production of human alpha-interferon in silkworm using a baculovirus vector. Nature 315:592–594CrossRefGoogle Scholar
  21. Motohashi T, Shimojima T, Fukagawa T, Maenaka K, Park EY (2005) Efficient large-scale protein production of larvae and pupae of silkworm by Bombyx mori nuclear polyhedrosis virus bacmid system. Biochem Biophys Res Commun 326:564–569. CrossRefGoogle Scholar
  22. Nguyen DH, Taub DD (2004) Targeting lipids to prevent HIV infection. Mol Interv 4:318–320. CrossRefGoogle Scholar
  23. Pitha J, Irie T, Sklar PB, Nye JS (1988) Drug solubilizers to aid pharmacologists: amorphous cyclodextrin derivatives. Life Sci 43:493–502CrossRefGoogle Scholar
  24. Rohrmann GF (2013) Baculovirus molecular biology. National Center for Biotechnology Information, Bethesda (MD)Google Scholar
  25. Shan L, Wang L, Yin J, Zhong P, Zhong J (2006) An OriP/EBNA-1-based baculovirus vector with prolonged and enhanced transgene expression. J Gene Med 8:1400–1406. CrossRefGoogle Scholar
  26. Yancey PG, Rodrigueza WV, Kilsdonk EP, Stoudt GW, Johnson WJ, Phillips MC, Rothblat GH (1996) Cellular cholesterol efflux mediated by cyclodextrins. Demonstration of kinetic pools and mechanism of efflux. J Biol Chem 271:16026–16034. CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Jinshan Huang
    • 1
    • 2
  • Na Liu
    • 1
  • Xingjia Shen
    • 1
    • 2
  • Bifang Hao
    • 1
    • 2
    Email author
  1. 1.Jiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of BiotechnologyJiangsu University of Science and TechnologyZhenjiangPeople’s Republic of China
  2. 2.Key Laboratory of Genetic Improvement of Sericulture in the Ministry of Agriculture, Sericultural Research InstituteChinese Academy of Agricultural ScienceZhenjiangPeople’s Republic of China

Personalised recommendations