Medium-throughput production of recombinant human proteins: protein production in insect cells

Methods Mol Biol. 2014:1091:95-121. doi: 10.1007/978-1-62703-691-7_6.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Batch Cell Culture Techniques
  • Cell Culture Techniques
  • Gene Expression*
  • Genetic Vectors / genetics
  • Humans
  • Proteins / genetics*
  • Proteins / metabolism*
  • Quality Control
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics*
  • Recombinant Proteins / isolation & purification
  • Sf9 Cells
  • Transfection

Substances

  • Proteins
  • Recombinant Proteins