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Metal-chelate affinity precipitation of proteins using responsive polymers

Mattiasson, Bo LU ; Kumar, Ashok LU ; Ivanov, Alexander LU and Galaev, Igor LU (2007) In Nature Protocols 2(1). p.213-220
Abstract
Affinity precipitation of proteins uses polymers capable of reversible soluble-insoluble transitions in response to small

environmental changes (temperature, pH or solvent composition). Here we describe protocols for (i) the synthesis of

responsive polymers with specific affinity to target proteins and (ii) the purification of proteins using these polymers.

The purification is based on precipitation of the affinity complex between the protein and the polymer, which is induced

by environmental changes. This separation strategy is simpler and more cost effective than conventional affinity column

chromatography. Specifically, we describe the synthesis of thermoresponsive... (More)
Affinity precipitation of proteins uses polymers capable of reversible soluble-insoluble transitions in response to small

environmental changes (temperature, pH or solvent composition). Here we describe protocols for (i) the synthesis of

responsive polymers with specific affinity to target proteins and (ii) the purification of proteins using these polymers.

The purification is based on precipitation of the affinity complex between the protein and the polymer, which is induced

by environmental changes. This separation strategy is simpler and more cost effective than conventional affinity column

chromatography. Specifically, we describe the synthesis of thermoresponsive 1-vinylimidazole:N-isopropylacrylamide

copolymers. The whole procedure takes 2–3 h when applied to purification of recombinant His-tag proteins or proteins with

natural metal binding groups by means of metal chelate affinity precipitation. Optimization of the polymer composition and

the type of chelating ions allows for target protein yields of 80% and higher. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Nature Protocols
volume
2
issue
1
pages
213 - 220
publisher
Nature Publishing Group
external identifiers
  • wos:000253138000028
  • scopus:34347255758
ISSN
1750-2799
DOI
10.1038/nprot.2006.440
language
English
LU publication?
yes
id
47457705-8623-4f79-8be7-177712190363 (old id 792693)
date added to LUP
2008-01-07 09:50:13
date last changed
2017-04-23 03:26:56
@article{47457705-8623-4f79-8be7-177712190363,
  abstract     = {Affinity precipitation of proteins uses polymers capable of reversible soluble-insoluble transitions in response to small<br/><br>
environmental changes (temperature, pH or solvent composition). Here we describe protocols for (i) the synthesis of<br/><br>
responsive polymers with specific affinity to target proteins and (ii) the purification of proteins using these polymers.<br/><br>
The purification is based on precipitation of the affinity complex between the protein and the polymer, which is induced<br/><br>
by environmental changes. This separation strategy is simpler and more cost effective than conventional affinity column<br/><br>
chromatography. Specifically, we describe the synthesis of thermoresponsive 1-vinylimidazole:N-isopropylacrylamide<br/><br>
copolymers. The whole procedure takes 2–3 h when applied to purification of recombinant His-tag proteins or proteins with<br/><br>
natural metal binding groups by means of metal chelate affinity precipitation. Optimization of the polymer composition and<br/><br>
the type of chelating ions allows for target protein yields of 80% and higher.},
  author       = {Mattiasson, Bo and Kumar, Ashok and Ivanov, Alexander and Galaev, Igor},
  issn         = {1750-2799},
  language     = {eng},
  number       = {1},
  pages        = {213--220},
  publisher    = {Nature Publishing Group},
  series       = {Nature Protocols},
  title        = {Metal-chelate affinity precipitation of proteins using responsive polymers},
  url          = {http://dx.doi.org/10.1038/nprot.2006.440},
  volume       = {2},
  year         = {2007},
}