A high-throughput mass spectrometry-based assay for large-scale profiling of circulating human apolipoproteins.

Autor:	Blanchard V; Université de La Réunion, INSERM, UMR 1188 Diabète athérothrombose Réunion Océan Indien (DéTROI), Plateforme CYROI, Saint-Denis de La Réunion, France. Electronic address: mailto:mikael.croyal@univ-nantes.fr., Garçon D; L'Institut du Thorax, INSERM, CNRS, University of Nantes, Nantes, France., Jaunet C; Department of Biochemistry, CHU de Nantes, France., Chemello K; Université de La Réunion, INSERM, UMR 1188 Diabète athérothrombose Réunion Océan Indien (DéTROI), Plateforme CYROI, Saint-Denis de La Réunion, France., Billon-Crossouard S; NUN, INRA, CHU Nantes, UMR 1280, PhAN, IMAD, CRNH-O, Nantes, France; CRNH-O Mass Spectrometry Core Facility, Nantes, France., Aguesse A; NUN, INRA, CHU Nantes, UMR 1280, PhAN, IMAD, CRNH-O, Nantes, France; CRNH-O Mass Spectrometry Core Facility, Nantes, France., Garfa A; CRNH-O Mass Spectrometry Core Facility, Nantes, France., Famchon G; Department of Biochemistry, CHU de Nantes, France., Torres A; NUN, INRA, CHU Nantes, UMR 1280, PhAN, IMAD, CRNH-O, Nantes, France., Le May C; L'Institut du Thorax, INSERM, CNRS, University of Nantes, Nantes, France., Pichelin M; L'Institut du Thorax, INSERM, CNRS, University of Nantes, CHU Nantes, Nantes, France., Bigot-Corbel E; Department of Biochemistry, CHU de Nantes, France., Lambert G; Université de La Réunion, INSERM, UMR 1188 Diabète athérothrombose Réunion Océan Indien (DéTROI), Plateforme CYROI, Saint-Denis de La Réunion, France., Cariou B; L'Institut du Thorax, INSERM, CNRS, University of Nantes, CHU Nantes, Nantes, France., Hadjadj S; CRNH-O Mass Spectrometry Core Facility, Nantes, France; L'Institut du Thorax, INSERM, CNRS, University of Nantes, CHU Nantes, Nantes, France., Krempf M; NUN, INRA, CHU Nantes, UMR 1280, PhAN, IMAD, CRNH-O, Nantes, France; CRNH-O Mass Spectrometry Core Facility, Nantes, France; ELSAN, Clinique Bretéché, Nantes, France., Bach-Ngohou K; Department of Biochemistry, CHU de Nantes, France; INSERM U1235, University of Nantes, Nantes, France., Croyal M; NUN, INRA, CHU Nantes, UMR 1280, PhAN, IMAD, CRNH-O, Nantes, France; CRNH-O Mass Spectrometry Core Facility, Nantes, France.
Jazyk:	angličtina
Zdroj:	Journal of lipid research [J Lipid Res] 2020 Jul; Vol. 61 (7), pp. 1128-1139. Date of Electronic Publication: 2020 May 13.
DOI:	10.1194/jlr.D120000835
Abstrakt:	Apolipoproteins govern lipoprotein metabolism and are promising biomarkers of metabolic and cardiovascular diseases. Unlike immunoassays, MS enables the quantification and phenotyping of multiple apolipoproteins. Hence, here, we aimed to develop a LC-MS/MS assay that can simultaneously quantitate 18 human apolipoproteins [A-I, A-II, A-IV, A-V, B48, B100, C-I, C-II, C-III, C-IV, D, E, F, H, J, L1, M, and (a)] and determined apoE, apoL1, and apo(a) phenotypes in human plasma and serum samples. The plasma and serum apolipoproteins were trypsin digested through an optimized procedure and peptides were extracted and analyzed by LC-MS/MS. The method was validated according to standard guidelines in samples spiked with known peptide amounts. The LC-MS/MS results were compared with those obtained with other techniques, and reproducibility, dilution effects, and stabilities were also assessed. Peptide markers were successfully selected for targeted apolipoprotein quantification and phenotyping. After optimization, the assay was validated for linearity, lower limits of quantification, accuracy (biases: -14.8% to 12.1%), intra-assay variability [coefficients of variation (CVs): 1.5-14.2%], and inter-assay repeatability (CVs: 4.1-14.3%). Bland-Altman plots indicated no major statistically significant differences between LC-MS/MS and other techniques. The LC-MS/MS results were reproducible over five repeated experiments (CVs: 1.8-13.7%), and we identified marked differences among the plasma and serum samples. The LC-MS/MS assay developed here is rapid, requires only small sampling volumes, and incurs reasonable costs, thus making it amenable for a wide range of studies of apolipoprotein metabolism. We also highlight how this assay can be implemented in laboratories. (Copyright © 2020 Blanchard et al.)
Databáze:	MEDLINE
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