Metal-chelate affinity precipitation of proteins using responsive polymers
(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:
https://lup.lub.lu.se/record/792693
- author
- Mattiasson, Bo LU ; Kumar, Ashok LU ; Ivanov, Alexander LU and Galaev, Igor LU
- organization
- publishing date
- 2007
- 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
- pmid:17401356
- 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
- 2016-04-01 11:39:43
- date last changed
- 2023-08-15 13:30:54
@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}},
doi = {{10.1038/nprot.2006.440}},
volume = {{2}},
year = {{2007}},
}