Purify First: Rapid expression and purification of proteins from XMRV

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 a...

Full description

Saved in:
Bibliographic Details
Published in:Protein expression and purification 2011-04, Vol.76 (2), p.238-247
Main Authors: Gillette, William K., Esposito, Dominic, Taylor, Troy E., Hopkins, Ralph F., Bagni, Rachel K., Hartley, James L.
Format: Article
Language:English
Subjects:
Animals
Baculoviridae - genetics
Base Sequence
Cloning, Molecular - methods
Electrophoresis, Polyacrylamide Gel
Escherichia coli - genetics
Molecular Sequence Data
Parallel purification
Protein Engineering - methods
Protein purification
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Small scale purification
Spodoptera
Xenotropic murine leukemia virus-related virus - chemistry
Xenotropic murine leukemia virus-related virus - genetics
Xenotropic murine leukemia virus-related virus - metabolism
XMRV
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2010.12.003