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Stolt-Bergner, P.* ; Benda, C.* ; Bergbrede, T.* ; Besir, H.* ; Celie, P.H.N.* ; Chang, C.J.* ; Drechsel, D.* ; Fischer, A.* ; Geerlof, A. ; Giabbai, B.* ; van den Heuvel, J.* ; Huber, G. ; Knecht, W.* ; Lehner, A.* ; Lemaitre, R.* ; Nordén, K.* ; Pardee, G.* ; Racke, I.* ; Remans, K.* ; Sander, A.* ; Scholz, J.* ; Stadnik, M.* ; Storici, P.* ; Weinbruch, D.* ; Zaror, I.* ; Lua, L.H.L.* ; Suppmann, S.*

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

J. Struct. Biol. 203, 71-80 (2018)
Postprint DOI PMC
Open Access Green
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((R)))or homologous recombination transfection within insect cells (e.g. flashBAC (TM)). 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, 204kDa; human ABL1 wildtype, 126kDa; human FMRP, 68 kDa; viral vNSl-Hl, 76kDa). 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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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Korrespondenzautor
Schlagwörter Baculovirus-insect Cell System ; Benchmark; Chitinase; Vectors; Vaccine; System
ISSN (print) / ISBN 1047-8477
e-ISSN 1047-8477
Zeitschrift Journal of Structural Biology
Quellenangaben Band: 203, Heft: 2, Seiten: 71-80 Artikelnummer: , Supplement: ,
Verlag Elsevier
Verlagsort 525 B St, Ste 1900, San Diego, Ca 92101-4495 Usa
Nichtpatentliteratur Publikationen
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Structural Biology (STB)