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This chapter describes the step-by-step methods employed by the Structural Genomics Consortium (SGC) for screening and producing proteins in the BacMam system. This eukaryotic expression system was selected and a screening process established in 2016 to enable production of highly challenging human integral membrane proteins (IMPs), which are a significant component of our target list. Here, we discuss our recently developed platform for identifying expression and monodispersity of IMPs from 3 mL of HEK293 cells.

Original publication

DOI

10.1007/978-1-0716-0892-0_6

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2021

Volume

2199

Pages

95 - 115

Keywords

BacMam, Baculovirus, FSEC, Gel filtration, IMAC, Insect cells, Mammalian cells, Protein purification, SEC chromatography, Transduction, Gene Expression, Genetic Vectors, HEK293 Cells, Humans, Membrane Proteins, Recombinant Proteins