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Adsorption of low-density lipoprotein, its oxidation, and subsequent binding of specific recombinant antibodies-an in situ ellipsometric study.

Stollenwerk, Maria ; Svensson, Olof ; Schiopu, Alexandru LU orcid ; Jansson, Bo LU ; Arnebrant, Thomas and Nordin Fredrikson, Gunilla LU (2011) In Biochimica et Biophysica Acta 1810(2). p.211-217
Abstract
BACKGROUND: Low-density lipoprotein (LDL) particles accumulate in the arterial wall and become oxidized during atherogenesis, leading to the formation of atherosclerotic plaques. The major protein of the LDL particle, apolipoprotein B-100 (apoB-100), becomes fragmented during oxidation and a target for the immune system. METHODS: In this study we used in situ ellipsometry to monitor the adsorption of LDL to solid silica surfaces and the effects of oxidation on the structure of the adsorbed LDL layer. We additionally investigated the binding kinetics of two recombinant human antibodies with different specificities recognizing epitopes of apoB-100 in surface-bound native and CuCl(2)-oxidized LDL (oxLDL). The latter process was studied by... (More)
BACKGROUND: Low-density lipoprotein (LDL) particles accumulate in the arterial wall and become oxidized during atherogenesis, leading to the formation of atherosclerotic plaques. The major protein of the LDL particle, apolipoprotein B-100 (apoB-100), becomes fragmented during oxidation and a target for the immune system. METHODS: In this study we used in situ ellipsometry to monitor the adsorption of LDL to solid silica surfaces and the effects of oxidation on the structure of the adsorbed LDL layer. We additionally investigated the binding kinetics of two recombinant human antibodies with different specificities recognizing epitopes of apoB-100 in surface-bound native and CuCl(2)-oxidized LDL (oxLDL). The latter process was studied by adsorbing LDL and then adding the antibody and CuCl(2) while continuously monitoring the amount of LDL adsorbed and the thickness of the film. The molar ratios between the antibodies and surface-bound LDL and oxLDL were calculated from these data. RESULTS: Our results indicate that oxidation of surface-bound LDL induces swelling of the layer, accompanied by a slight desorption. We further found that both antibodies were able to recognize LDL and oxLDL in its adsorbed orientation. Quantitative information was obtained on the number of available binding sites on surface-bound LDL and oxLDL for these two antibodies. GENERAL SIGNIFICANCE: Using ellipsometry for real-time monitoring of adsorption, in situ oxidation of LDL and binding of specific recombinant antibodies to surface-bound LDL, will open up possibilities to map different conformations and orientations of LDL in the adsorbed state. (Less)
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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Antibody, Adsorption, apoB-100, Ellipsometry, Oxidation, Low-density, lipoprotein (LDL)
in
Biochimica et Biophysica Acta
volume
1810
issue
2
pages
211 - 217
publisher
Elsevier
external identifiers
  • wos:000286543100009
  • pmid:20970483
  • scopus:78650677105
  • pmid:20970483
ISSN
0006-3002
DOI
10.1016/j.bbagen.2010.10.006
language
English
LU publication?
yes
id
a5f2dbb2-726b-4598-875c-64fad546cc51 (old id 1710836)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/20970483?dopt=Abstract
date added to LUP
2016-04-01 14:43:51
date last changed
2024-10-10 21:00:21
@article{a5f2dbb2-726b-4598-875c-64fad546cc51,
  abstract     = {{BACKGROUND: Low-density lipoprotein (LDL) particles accumulate in the arterial wall and become oxidized during atherogenesis, leading to the formation of atherosclerotic plaques. The major protein of the LDL particle, apolipoprotein B-100 (apoB-100), becomes fragmented during oxidation and a target for the immune system. METHODS: In this study we used in situ ellipsometry to monitor the adsorption of LDL to solid silica surfaces and the effects of oxidation on the structure of the adsorbed LDL layer. We additionally investigated the binding kinetics of two recombinant human antibodies with different specificities recognizing epitopes of apoB-100 in surface-bound native and CuCl(2)-oxidized LDL (oxLDL). The latter process was studied by adsorbing LDL and then adding the antibody and CuCl(2) while continuously monitoring the amount of LDL adsorbed and the thickness of the film. The molar ratios between the antibodies and surface-bound LDL and oxLDL were calculated from these data. RESULTS: Our results indicate that oxidation of surface-bound LDL induces swelling of the layer, accompanied by a slight desorption. We further found that both antibodies were able to recognize LDL and oxLDL in its adsorbed orientation. Quantitative information was obtained on the number of available binding sites on surface-bound LDL and oxLDL for these two antibodies. GENERAL SIGNIFICANCE: Using ellipsometry for real-time monitoring of adsorption, in situ oxidation of LDL and binding of specific recombinant antibodies to surface-bound LDL, will open up possibilities to map different conformations and orientations of LDL in the adsorbed state.}},
  author       = {{Stollenwerk, Maria and Svensson, Olof and Schiopu, Alexandru and Jansson, Bo and Arnebrant, Thomas and Nordin Fredrikson, Gunilla}},
  issn         = {{0006-3002}},
  keywords     = {{Antibody; Adsorption; apoB-100; Ellipsometry; Oxidation; Low-density; lipoprotein (LDL)}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{211--217}},
  publisher    = {{Elsevier}},
  series       = {{Biochimica et Biophysica Acta}},
  title        = {{Adsorption of low-density lipoprotein, its oxidation, and subsequent binding of specific recombinant antibodies-an in situ ellipsometric study.}},
  url          = {{http://dx.doi.org/10.1016/j.bbagen.2010.10.006}},
  doi          = {{10.1016/j.bbagen.2010.10.006}},
  volume       = {{1810}},
  year         = {{2011}},
}