E. coli and insect cell expression, automated purification and quantitative analysis.
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.
Chambers
Stephen P
SP
Gene Expression, Vertex Pharmaceuticals, Cambridge, MA, USA.
Fulghum
John R
JR
Austen
Douglas A
DA
Lu
Fan
F
Swalley
Susanne E
SE
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*
D011994:Recombinant Proteins