Improved Non-chromatographic Purification of a Recombinant Protein by Cationic Elastin-like Polypeptides

This paper reports an improvement in the purification of thioredoxin (Trx) expressed from E. coli by inverse transition cycling (ITC) using cationic elastin-like polypeptides (ELPs). Two ELP libraries having 2% and 5% lysine residues and molecular weights ranging from 4 to 61.1 kDa showed greater sa...

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Bibliographic Details
Published in:Biomacromolecules 2007-05, Vol.8 (5), p.1417-1424
Main Authors: Lim, Dong Woo, Trabbic-Carlson, Kimberly, MacKay, J. Andrew, Chilkoti, Ashutosh
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Sequence
Biological and medical sciences
Biotechnology
Cations - chemistry
Elastin - biosynthesis
Elastin - chemistry
Elastin - genetics
Escherichia coli - chemistry
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Methods. Procedures. Technologies
Molecular Sequence Data
Others
Peptide Library
Peptides - chemistry
Peptides - genetics
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - isolation & purification
Sodium Chloride - chemistry
Thioredoxins - biosynthesis
Thioredoxins - chemistry
Thioredoxins - isolation & purification
Various methods and equipments
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Summary:This paper reports an improvement in the purification of thioredoxin (Trx) expressed from E. coli by inverse transition cycling (ITC) using cationic elastin-like polypeptides (ELPs). Two ELP libraries having 2% and 5% lysine residues and molecular weights ranging from 4 to 61.1 kDa showed greater salt sensitivity in their inverse transition behavior than purely aliphatic ELPs. Expression yield of Trx-ELP fusions was an unpredictable function of guest residue composition, but reducing the molecular weight of the ELP tag generally increased Trx yield. A cationic 4.3 kDa ELP is the shortest ELP used to purify any protein by ITC to date. A 15.9 kDa ELP with a guest residue composition of K:V:F of 1:7:1 was found to be the optimal cationic tag to purify Trx, as it provided 50% greater Trx yield and only required one-fifth the added NaCl for purification of Trx as compared to previously used aliphatic ELP tags.
ISSN:1525-7797
1526-4602
DOI:10.1021/bm060849t