Purification of proteins from baculovirus-infected insect cells.
Expression of recombinant proteins in the baculovirus/insect cell expression system is employed because it enables post-translational protein modification and high yields of recombinant protein. The system is capable of facilitating the functional expression of many proteins - either secreted or intracellularly located within infected insect cells. Strategies for the isolation and extraction of soluble proteins are presented in this chapter and involve selective cell lysis, precipitation and chromatography. Protein insolubility, following recombinant expression in insect cells, can occur. However, using the methods described herein, it is possible to extract and purify insoluble protein using affinity, ion-exchange and gel filtration chromatography. Indeed, protein insolubility often aids protein purification.
O'Shaughnessy
Luke
L
Department of Biology, National Institute for Cellular Biotechnology, National University of Ireland Maynooth, Maynooth, Co. Kildare, Ireland.
Doyle
Sean
S
D000645:Ammonium Sulfate / Q000737:chemistry; D053000:Analytic Sample Preparation Methods; D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000502:physiology*; D005591:Chemical Fractionation / Q000379:methods*; D011232:Chemical Precipitation; D002845:Chromatography; D007313:Insecta / Q000166:cytology* / Q000235:genetics / Q000821:virology*; D011489:Protein Denaturation; D015202:Protein Engineering / Q000379:methods*; D021381:Protein Transport; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
D011994:Recombinant Proteins; D000645:Ammonium Sulfate