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A new approach to examine conformational changes occurring upon binding of ligand by biomolecules

Wingren, Christer LU and Hansson, Ulla-Britt (1995) In Biochimica et Biophysica Acta. General Subjects 1244(1). p.209-215
Abstract
Liquid-liquid partition chromatography in an aqueous poly(ethylene glycol)/dextran two-phase system (LLPC) is shown to be a quick and sensitive method for detecting conformational changes occurring upon binding of ligands by biospecific molecules. Two groups of well-characterized proteins, enzymes and monoclonal antibodies, were employed. As an example, LLPC demonstrated that isoforms of lactate dehydrogenase as well as of hexokinase existed in a ligand-dependent equilibrium between two forms and that conformational changes occurred when monoclonal antibodies bound haptens. We also demonstrate that the method could be used to detect and separate subfractions in preparations of unliganded proteins that appeared to be homogeneous when... (More)
Liquid-liquid partition chromatography in an aqueous poly(ethylene glycol)/dextran two-phase system (LLPC) is shown to be a quick and sensitive method for detecting conformational changes occurring upon binding of ligands by biospecific molecules. Two groups of well-characterized proteins, enzymes and monoclonal antibodies, were employed. As an example, LLPC demonstrated that isoforms of lactate dehydrogenase as well as of hexokinase existed in a ligand-dependent equilibrium between two forms and that conformational changes occurred when monoclonal antibodies bound haptens. We also demonstrate that the method could be used to detect and separate subfractions in preparations of unliganded proteins that appeared to be homogeneous when analysed by other techniques. (Less)
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author
and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Conformational change, Phase partition, LLPC, Ligand binding, Antibody, Enzyme
in
Biochimica et Biophysica Acta. General Subjects
volume
1244
issue
1
pages
209 - 215
publisher
Elsevier
external identifiers
  • scopus:0029011518
ISSN
0304-4165
DOI
10.1016/0304-4165(95)00007-X
language
English
LU publication?
yes
id
f5041780-d8f9-4f58-8e8c-e26b447eb643 (old id 126508)
date added to LUP
2016-04-01 16:12:25
date last changed
2021-01-03 10:09:11
@article{f5041780-d8f9-4f58-8e8c-e26b447eb643,
  abstract     = {{Liquid-liquid partition chromatography in an aqueous poly(ethylene glycol)/dextran two-phase system (LLPC) is shown to be a quick and sensitive method for detecting conformational changes occurring upon binding of ligands by biospecific molecules. Two groups of well-characterized proteins, enzymes and monoclonal antibodies, were employed. As an example, LLPC demonstrated that isoforms of lactate dehydrogenase as well as of hexokinase existed in a ligand-dependent equilibrium between two forms and that conformational changes occurred when monoclonal antibodies bound haptens. We also demonstrate that the method could be used to detect and separate subfractions in preparations of unliganded proteins that appeared to be homogeneous when analysed by other techniques.}},
  author       = {{Wingren, Christer and Hansson, Ulla-Britt}},
  issn         = {{0304-4165}},
  keywords     = {{Conformational change; Phase partition; LLPC; Ligand binding; Antibody; Enzyme}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{209--215}},
  publisher    = {{Elsevier}},
  series       = {{Biochimica et Biophysica Acta. General Subjects}},
  title        = {{A new approach to examine conformational changes occurring upon binding of ligand by biomolecules}},
  url          = {{http://dx.doi.org/10.1016/0304-4165(95)00007-X}},
  doi          = {{10.1016/0304-4165(95)00007-X}},
  volume       = {{1244}},
  year         = {{1995}},
}