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Assessment of Peptide Chemical Modifications on the Development of an Accurate and Precise Multiplex Selected Reaction Monitoring Assay for Apolipoprotein E Isoforms

Martinez-Morillo, Eduardo; Nielsen, Henrietta LU ; Batruch, Ihor; Drabovich, Andrei P.; Begcevic, Ilijana; Lopez, Mary F.; Minthon, Lennart LU ; Bu, Guojun; Mattsson, Niklas and Portelius, Erik, et al. (2014) In Journal of Proteome Research 13(2). p.1077-1087
Abstract
Apolipoprotein E (ApoE) is a polymorphic protein that plays a major role in lipid metabolism in the central nervous system and periphery. It has three common allelic isoforms, ApoE2, ApoE3 and ApoE4, that differ in only one or two amino acids. ApoE isoforms have been associated with the occurence and progression of several pathological conditions, such as coronary atherosclerosis and Alzheimer's disease. The aim of this study was to develop a mass spectrometry (MS)-based assay for absolute quantification of ApoE isoforms in cerebrospinal fluid and plasma samples using isotope-labeled peptides. The assay included five tryptic peptides: CLAVYQAGAR (ApoE2), LGADMEDVCGR (ApoE2 and 3);. LAVYQAGAR (ApoE3 and 4), LGAD-MEDVR (ApoE4), and LGPLVEQGR... (More)
Apolipoprotein E (ApoE) is a polymorphic protein that plays a major role in lipid metabolism in the central nervous system and periphery. It has three common allelic isoforms, ApoE2, ApoE3 and ApoE4, that differ in only one or two amino acids. ApoE isoforms have been associated with the occurence and progression of several pathological conditions, such as coronary atherosclerosis and Alzheimer's disease. The aim of this study was to develop a mass spectrometry (MS)-based assay for absolute quantification of ApoE isoforms in cerebrospinal fluid and plasma samples using isotope-labeled peptides. The assay included five tryptic peptides: CLAVYQAGAR (ApoE2), LGADMEDVCGR (ApoE2 and 3);. LAVYQAGAR (ApoE3 and 4), LGAD-MEDVR (ApoE4), and LGPLVEQGR (total ApoE). Both cerebrospinal fluid and plasma samples were assayed to validate the method. The digestion yield and the extension of chemical modifications in selected amino acid residues (methionine oxidation, glutamine deamidation, and cyclization of N-terminus carbamidomethylcysteine) were also studied. The ApoE phenotype was successfully assigned to all samples analyzed in a blinded manner. The method showed good linearity (R-2 > 0.99) and reproducibility (within laboratory imprecision <13%). The comparison of the MS-based assay with an ELISA for total ApoE concentration showed a moderate correlation (R-2 = 0.59). This MS-based assay can serve as an important tool in clinical studies aiming to elucidate the association between ApoE genotype, total ApoE, and ApoE isoform concentrations in various disorders related to ApoE polymorphisms. (Less)
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Contribution to journal
publication status
published
subject
keywords
apolipoprotein E, cerebrospinal fluid, chemical modification, isoform, peptide, plasma, selected reaction monitoring
in
Journal of Proteome Research
volume
13
issue
2
pages
1077 - 1087
publisher
The American Chemical Society
external identifiers
  • wos:000331164100066
  • scopus:84893826356
ISSN
1535-3893
DOI
10.1021/pr401060x
language
English
LU publication?
yes
id
eca83a47-ef9b-4d0f-8047-bd9c95fce2eb (old id 4368423)
date added to LUP
2014-05-05 08:36:26
date last changed
2017-04-09 03:13:37
@article{eca83a47-ef9b-4d0f-8047-bd9c95fce2eb,
  abstract     = {Apolipoprotein E (ApoE) is a polymorphic protein that plays a major role in lipid metabolism in the central nervous system and periphery. It has three common allelic isoforms, ApoE2, ApoE3 and ApoE4, that differ in only one or two amino acids. ApoE isoforms have been associated with the occurence and progression of several pathological conditions, such as coronary atherosclerosis and Alzheimer's disease. The aim of this study was to develop a mass spectrometry (MS)-based assay for absolute quantification of ApoE isoforms in cerebrospinal fluid and plasma samples using isotope-labeled peptides. The assay included five tryptic peptides: CLAVYQAGAR (ApoE2), LGADMEDVCGR (ApoE2 and 3);. LAVYQAGAR (ApoE3 and 4), LGAD-MEDVR (ApoE4), and LGPLVEQGR (total ApoE). Both cerebrospinal fluid and plasma samples were assayed to validate the method. The digestion yield and the extension of chemical modifications in selected amino acid residues (methionine oxidation, glutamine deamidation, and cyclization of N-terminus carbamidomethylcysteine) were also studied. The ApoE phenotype was successfully assigned to all samples analyzed in a blinded manner. The method showed good linearity (R-2 &gt; 0.99) and reproducibility (within laboratory imprecision &lt;13%). The comparison of the MS-based assay with an ELISA for total ApoE concentration showed a moderate correlation (R-2 = 0.59). This MS-based assay can serve as an important tool in clinical studies aiming to elucidate the association between ApoE genotype, total ApoE, and ApoE isoform concentrations in various disorders related to ApoE polymorphisms.},
  author       = {Martinez-Morillo, Eduardo and Nielsen, Henrietta and Batruch, Ihor and Drabovich, Andrei P. and Begcevic, Ilijana and Lopez, Mary F. and Minthon, Lennart and Bu, Guojun and Mattsson, Niklas and Portelius, Erik and Hansson, Oskar and Diamandis, Eleftherios P.},
  issn         = {1535-3893},
  keyword      = {apolipoprotein E,cerebrospinal fluid,chemical modification,isoform,peptide,plasma,selected reaction monitoring},
  language     = {eng},
  number       = {2},
  pages        = {1077--1087},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {Assessment of Peptide Chemical Modifications on the Development of an Accurate and Precise Multiplex Selected Reaction Monitoring Assay for Apolipoprotein E Isoforms},
  url          = {http://dx.doi.org/10.1021/pr401060x},
  volume       = {13},
  year         = {2014},
}