title: E. coli and insect cell expression, automated purification and quantitative analysis.
abstract: The production of recombinant proteins usually involves the exploration of a wide variety of expression and purification methodologies in the pursuit of a strategy tailored to a particular protein. The methods applied are reliant on exploiting individual differences between expression systems or the variations in specific protein properties. These bespoke strategies have not lent themselves to high-throughput methodologies. Ultimately the development of robust generic methods capable of simplifying and stabilizing the process, allowing automation, was necessary to increase throughput. This chapter describes a series of high-throughput methods used to express, purify, and quantify recombinant protein produced in E. coli or insect cells.
authors.affiliation: Gene Expression, Vertex Pharmaceuticals, Cambridge, MA, USA.
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Chambers
authors.forename: John R
authors.initials: JR
authors.lastname: Fulghum
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Austen
authors.forename: Fan
authors.initials: F
authors.lastname: Lu
authors.forename: Susanne E
authors.initials: SE
authors.lastname: Swalley
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000254:growth & development; D002477:Cells / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D007313:Insecta / Q000235:genetics* / Q000821:virology; D046208:Microchip Analytical Procedures; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins