title: Purify First: rapid expression and purification of proteins from XMRV.
abstract: Purifying proteins from recombinant sources is often difficult, time-consuming, and costly. We have recently instituted a series of improvements in our protein purification pipeline that allows much more accurate choice of expression host and conditions and purification protocols. The key elements are parallel cloning, small scale parallel expression and lysate preparation, and small scale parallel protein purification. Compared to analyzing expression data only, results from multiple small scale protein purifications predict success at scale-up with greatly improved reliability. Using these new procedures we purified eight of nine proteins from xenotropic murine leukemia virus-related virus (XMRV) on the first attempt at large scale.
authors.affiliation: Protein Expression Laboratory, SAIC-Frederick Inc., NCI-Frederick, Frederick, MD 21702, USA.
authors.forename: William K
authors.initials: WK
authors.lastname: Gillette
authors.forename: Dominic
authors.initials: D
authors.lastname: Esposito
authors.forename: Troy E
authors.initials: TE
authors.lastname: Taylor
authors.forename: Ralph F
authors.initials: RF
authors.lastname: Hopkins
authors.forename: Rachel K
authors.initials: RK
authors.lastname: Bagni
authors.forename: James L
authors.initials: JL
authors.lastname: Hartley
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D018411:Spodoptera; D058008:Xenotropic murine leukemia virus-related virus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins