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Baculovirus-driven protein expression in insect cells : A benchmarking study

Stolt-Bergner, Peggy ; Benda, Christian ; Bergbrede, Tim ; Besir, Hüseyin ; Celie, Patrick H.N. ; Chang, Cindy ; Drechsel, David ; Fischer, Ariane ; Geerlof, Arie and Giabbai, Barbara , et al. (2018) In Journal of Structural Biology 203(2). p.71-80
Abstract

Baculovirus-insect cell expression system has become one of the most widely used eukaryotic expression systems for heterologous protein production in many laboratories. The availability of robust insect cell lines, serum-free media, a range of vectors and commercially-packaged kits have supported the demand for maximizing the exploitation of the baculovirus-insect cell expression system. Naturally, this resulted in varied strategies adopted by different laboratories to optimize protein production. Most laboratories have preference in using either the E. coli transposition-based recombination bacmid technology (e.g. Bac-to-Bac®) or homologous recombination transfection within insect cells (e.g. flashBAC™). Limited data is presented in... (More)

Baculovirus-insect cell expression system has become one of the most widely used eukaryotic expression systems for heterologous protein production in many laboratories. The availability of robust insect cell lines, serum-free media, a range of vectors and commercially-packaged kits have supported the demand for maximizing the exploitation of the baculovirus-insect cell expression system. Naturally, this resulted in varied strategies adopted by different laboratories to optimize protein production. Most laboratories have preference in using either the E. coli transposition-based recombination bacmid technology (e.g. Bac-to-Bac®) or homologous recombination transfection within insect cells (e.g. flashBAC™). Limited data is presented in the literature to benchmark the protocols used for these baculovirus vectors to facilitate the selection of a system for optimal production of target proteins. Taking advantage of the Protein Production and Purification Partnership in Europe (P4EU) scientific network, a benchmarking initiative was designed to compare the diverse protocols established in thirteen individual laboratories. This benchmarking initiative compared the expression of four selected intracellular proteins (mouse Dicer-2, 204 kDa; human ABL1 wildtype, 126 kDa; human FMRP, 68 kDa; viral vNS1-H1, 76 kDa). Here, we present the expression and purification results on these proteins and highlight the significant differences in expression yields obtained using different commercially-packaged baculovirus vectors. The highest expression level for difficult-to-express intracellular protein candidates were observed with the EmBacY baculovirus vector system.

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subject
keywords
Baculovirus-insect cell system, Benchmark
in
Journal of Structural Biology
volume
203
issue
2
pages
71 - 80
publisher
Elsevier
external identifiers
  • pmid:29545204
  • scopus:85044317706
ISSN
1047-8477
DOI
10.1016/j.jsb.2018.03.004
language
English
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yes
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2eb373f5-95fb-4a76-9f51-e19beca50b20
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2018-04-10 07:49:06
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2020-07-08 04:15:22
@article{2eb373f5-95fb-4a76-9f51-e19beca50b20,
  abstract     = {<p>Baculovirus-insect cell expression system has become one of the most widely used eukaryotic expression systems for heterologous protein production in many laboratories. The availability of robust insect cell lines, serum-free media, a range of vectors and commercially-packaged kits have supported the demand for maximizing the exploitation of the baculovirus-insect cell expression system. Naturally, this resulted in varied strategies adopted by different laboratories to optimize protein production. Most laboratories have preference in using either the E. coli transposition-based recombination bacmid technology (e.g. Bac-to-Bac®) or homologous recombination transfection within insect cells (e.g. flashBAC™). Limited data is presented in the literature to benchmark the protocols used for these baculovirus vectors to facilitate the selection of a system for optimal production of target proteins. Taking advantage of the Protein Production and Purification Partnership in Europe (P4EU) scientific network, a benchmarking initiative was designed to compare the diverse protocols established in thirteen individual laboratories. This benchmarking initiative compared the expression of four selected intracellular proteins (mouse Dicer-2, 204 kDa; human ABL1 wildtype, 126 kDa; human FMRP, 68 kDa; viral vNS1-H1, 76 kDa). Here, we present the expression and purification results on these proteins and highlight the significant differences in expression yields obtained using different commercially-packaged baculovirus vectors. The highest expression level for difficult-to-express intracellular protein candidates were observed with the EmBacY baculovirus vector system.</p>},
  author       = {Stolt-Bergner, Peggy and Benda, Christian and Bergbrede, Tim and Besir, Hüseyin and Celie, Patrick H.N. and Chang, Cindy and Drechsel, David and Fischer, Ariane and Geerlof, Arie and Giabbai, Barbara and van den Heuvel, Joop and Huber, Georg and Knecht, Wolfgang and Lehner, Anita and Lemaitre, Regis and Nordén, Kristina and Pardee, Gwynn and Racke, Ines and Remans, Kim and Sander, Astrid and Scholz, Judith and Stadnik, Magda and Storici, Paola and Weinbruch, Daniel and Zaror, Isabel and Lua, Linda H.L. and Suppmann, Sabine},
  issn         = {1047-8477},
  language     = {eng},
  month        = {03},
  number       = {2},
  pages        = {71--80},
  publisher    = {Elsevier},
  series       = {Journal of Structural Biology},
  title        = {Baculovirus-driven protein expression in insect cells : A benchmarking study},
  url          = {http://dx.doi.org/10.1016/j.jsb.2018.03.004},
  doi          = {10.1016/j.jsb.2018.03.004},
  volume       = {203},
  year         = {2018},
}