title: Medium-throughput production of recombinant human proteins: protein production in insect cells.
abstract: This chapter describes the step-by-step methods employed by the Structural Genomics Consortium (SGC) for screening and producing proteins in the baculovirus expression vector system (BEVS). This eukaryotic expression system was selected and a screening process established in 2007 as a measure to tackle the more challenging kinase, RNA-DNA processing and integral membrane protein families on our target list. Here, we discuss our platform for identifying soluble proteins from 3 ml of insect cell culture and describe the procedures involved in producing protein from liter-scale cultures. Although not discussed in this chapter, the same process can also be applied to integral membrane proteins (IMPs) with slight adaptations to the purification procedure.
authors.affiliation: Structural Genomics Consortium, University of Oxford, Oxford, UK.
authors.forename: Pravin
authors.initials: P
authors.lastname: Mahajan
authors.forename: Claire
authors.initials: C
authors.lastname: Strain-Damerell
authors.forename: Opher
authors.initials: O
authors.lastname: Gileadi
authors.forename: Nicola A
authors.initials: NA
authors.lastname: Burgess-Brown
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D061249:Batch Cell Culture Techniques; D018929:Cell Culture Techniques; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D011506:Proteins / Q000235:genetics* / Q000378:metabolism*; D011786:Quality Control; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D061987:Sf9 Cells; D014162:Transfection
chemical_list: D011506:Proteins; D011994:Recombinant Proteins