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Baculovirus-driven protein expression in insect cells : A benchmarking study
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- Stolt-Bergner, Peggy (author)
- Vienna Biocenter (VBC)
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- Benda, Christian (author)
- Max Planck Institute of Biochemistry
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- Bergbrede, Tim (author)
- Lead Discovery Center GmbH
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- Besir, Hüseyin (author)
- European Molecular Biology Laboratory Heidelberg
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- Celie, Patrick H.N. (author)
- Netherlands Cancer Institute
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- Chang, Cindy (author)
- University of Queensland
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- Drechsel, David (author)
- Max Planck Institute of Molecular Cell Biology and Genetics
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- Fischer, Ariane (author)
- Max Planck Institute of Biochemistry
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- Geerlof, Arie (author)
- Helmholtz Zentrum München
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- Giabbai, Barbara (author)
- Elettra Sincrotrone Trieste
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- van den Heuvel, Joop (author)
- Helmholtz Centre for Infection Research
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- Huber, Georg (author)
- Helmholtz Zentrum München
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- Knecht, Wolfgang (author)
- Lund University,Lunds universitet,Molekylär cellbiologi,Biologiska institutionen,Naturvetenskapliga fakulteten,Molecular Cell Biology,Department of Biology,Faculty of Science
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- Lehner, Anita (author)
- Vienna Biocenter (VBC)
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- Lemaitre, Regis (author)
- Max Planck Institute of Molecular Cell Biology and Genetics
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- Nordén, Kristina (author)
- Lund University,Lunds universitet,Medicinsk proteinvetenskap,Forskargrupper vid Lunds universitet,Medical Protein Science,Lund University Research Groups
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- Pardee, Gwynn (author)
- Novartis Institutes for BioMedical Research, Inc.
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- Racke, Ines (author)
- European Molecular Biology Laboratory Heidelberg
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- Remans, Kim (author)
- European Molecular Biology Laboratory Heidelberg
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- Sander, Astrid (author)
- Max-Planck-Institute for Molecular Plant Physiology
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- Scholz, Judith (author)
- Max Planck Institute of Biochemistry
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- Stadnik, Magda (author)
- Netherlands Cancer Institute
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- Storici, Paola (author)
- Elettra Sincrotrone Trieste
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- Weinbruch, Daniel (author)
- Coriolis Pharma
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- Zaror, Isabel (author)
- Novartis Institutes for BioMedical Research, Inc.
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- Lua, Linda H.L. (author)
- University of Queensland
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- Suppmann, Sabine (author)
- Max Planck Institute of Biochemistry
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(creator_code:org_t)
- Elsevier BV, 2018
- 2018
- English.
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In: Journal of Structural Biology. - : Elsevier BV. - 1047-8477. ; 203:2, s. 71-80
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Abstract
Subject headings
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- 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.
Subject headings
- NATURVETENSKAP -- Biologi -- Strukturbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Structural Biology (hsv//eng)
Keyword
- Baculovirus-insect cell system
- Benchmark
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- ref (subject category)
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- By the author/editor
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Stolt-Bergner, P ...
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Benda, Christian
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Bergbrede, Tim
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Besir, Hüseyin
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Celie, Patrick H ...
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Chang, Cindy
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show more...
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Drechsel, David
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Fischer, Ariane
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Geerlof, Arie
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Giabbai, Barbara
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van den Heuvel, ...
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Huber, Georg
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Knecht, Wolfgang
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Lehner, Anita
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Lemaitre, Regis
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Nordén, Kristina
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Pardee, Gwynn
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Racke, Ines
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Remans, Kim
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Sander, Astrid
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Scholz, Judith
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Stadnik, Magda
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Storici, Paola
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Weinbruch, Danie ...
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Zaror, Isabel
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Lua, Linda H.L.
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Suppmann, Sabine
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Structural Biolo ...
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Journal of Struc ...
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Lund University