SweetBac: Applying MultiBac Technology Towards Flexible Modification of Insect Cell Glycosylation

  • Dieter Palmberger
  • Dubravko Rendic
Part of the Methods in Molecular Biology book series (MIMB, volume 1321)


Observed different glycosylation patterns of glycoconjugates (recombinantly) produced in various eukaryotic organisms are a direct consequence of differences in numerous proteins involved in biosynthesis of the relevant glycan chains in these species. The need for efficient, robust and flexible methods for recombinant expression of proteins is met by the recently described MultiBac technology, an advanced and optimized baculovirus-based system for simultaneous recombinant protein expression in insect cells. A derivative of MultiBac technology, the SweetBac system aims at the modification of the glycosylation potential of insect cells as expression hosts. The application of SweetBac, including the methods needed to investigate the glycosylation pattern of the purified recombinant protein, is described in this chapter.

Key words

SweetBac MultiBac Baculovirus Insect cells Recombinant 


  1. 1.
    Shields RL, Lai J, Keck R et al (2002) Lack of fucose on human IgG1 N-linked oligosaccharide improves binding to human Fcgamma RIII and antibody-dependent cellular toxicity. J Biol Chem 277:26733–26740PubMedCrossRefGoogle Scholar
  2. 2.
    Kanda Y, Yamada T, Mori K et al (2007) Comparison of biological activity among nonfucosylated therapeutic IgG1 antibodies with three different N-linked Fc oligosaccharides: the high-mannose, hybrid, and complex types. Glycobiology 17:104–118PubMedCrossRefGoogle Scholar
  3. 3.
    Bieniossek C, Richmond TJ, Berger I (2008) MultiBac: multigene baculovirus-based eukaryotic protein complex production. Curr Protoc Protein Sci. Chapter 5; Unit 5: 20Google Scholar
  4. 4.
    Bieniossek C, Imasaki T, Takagi Y et al (2012) MultiBac: expanding the research toolbox for multiprotein complexes. Trends Biochem Sci 37:49–57PubMedCrossRefGoogle Scholar
  5. 5.
    Fitzgerald DJ, Berger P, Schaffitze C et al (2006) Protein complex expression by using multigene baculoviral vectors. Nat Methods 3:1021–1032PubMedCrossRefGoogle Scholar
  6. 6.
    Berger I, Garzoni F, Chaillet M et al (2013) The multiBac protein complex production platform at the EMBL. J Visual Exp (77):e50159. doi: 10.3791/50159
  7. 7.
    Rendic D, Wilson IBH, Paschinger K (2008) The glycosylation capacity of insect cells. Croatica Chem Acta 81:7–21Google Scholar
  8. 8.
    Rendic D, Wilson IBH, Lubec G et al (2007) Adaptation of the “in-gel release method” to N-glycome analysis of low-milligram amounts of material. Electrophoresis 28:4484–4492PubMedCrossRefGoogle Scholar
  9. 9.
    Palmberger D, Rendic D, Tauber P et al (2011) Insect cells for antibody production: evaluation of an efficient alternative. J Biotechnol 153:160–166PubMedCrossRefGoogle Scholar
  10. 10.
    Cox MM (2012) Recombinant protein vaccines produced in insect cells. Vaccine 30:1759–1766PubMedCrossRefGoogle Scholar
  11. 11.
    Palmberger D, Ashjaei K, Strell S et al (2014) Minimizing fucosylation in insect cell-derived glycoproteins reduces binding to IgE antibodies from the sera of patients with allergy. Biotechnol J. doi: 10.1002/biot.201300061 Google Scholar
  12. 12.
    Mabashi-Asazuma H, Kuo CW, Khoo KH et al (2014) A novel baculovirus vector for the production of nonfucosylated recombinant glycoproteins in insect cells. Glycobiology 24:325–340PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Palmberger D, Klausberger M, Berger I et al (2013) MultiBac turns sweet. Bioengineered 4:78–83PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Palmberger D, Wilson IBH, Berger I et al (2012) SweetBac: a new approach for the production of mammalianised glycoproteins in insect cells. PLoS One 7, e34226PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Krammer F, Margine I, Tan GS et al (2012) A carboxy-terminal trimerization domain stabilizes conformational epitopes on the stalk domain of soluble recombinant hemagglutinin substrates. PLoS One 7, e43603PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Dieter Palmberger
    • 1
  • Dubravko Rendic
    • 2
  1. 1.Department of BiotechnologyUniversity of Natural Resources and Life SciencesViennaAustria
  2. 2.Department of ChemistryUniversity of Natural Resources and Life SciencesViennaAustria

Personalised recommendations