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ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing
Despite intense interest, methods that provide enhanced sensitivity and specificity in parallel measurements of candidate protein biomarkers in numerous samples have been lacking. We present herein a multiplex proximity ligation assay with readout via realtime PCR or DNA sequencing (ProteinSeq). We...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3183061/ https://www.ncbi.nlm.nih.gov/pubmed/21980495 http://dx.doi.org/10.1371/journal.pone.0025583 |
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author | Darmanis, Spyros Nong, Rachel Yuan Vänelid, Johan Siegbahn, Agneta Ericsson, Olle Fredriksson, Simon Bäcklin, Christofer Gut, Marta Heath, Simon Gut, Ivo Glynne Wallentin, Lars Gustafsson, Mats G. Kamali-Moghaddam, Masood Landegren, Ulf |
author_facet | Darmanis, Spyros Nong, Rachel Yuan Vänelid, Johan Siegbahn, Agneta Ericsson, Olle Fredriksson, Simon Bäcklin, Christofer Gut, Marta Heath, Simon Gut, Ivo Glynne Wallentin, Lars Gustafsson, Mats G. Kamali-Moghaddam, Masood Landegren, Ulf |
author_sort | Darmanis, Spyros |
collection | PubMed |
description | Despite intense interest, methods that provide enhanced sensitivity and specificity in parallel measurements of candidate protein biomarkers in numerous samples have been lacking. We present herein a multiplex proximity ligation assay with readout via realtime PCR or DNA sequencing (ProteinSeq). We demonstrate improved sensitivity over conventional sandwich assays for simultaneous analysis of sets of 35 proteins in 5 µl of blood plasma. Importantly, we observe a minimal tendency to increased background with multiplexing, compared to a sandwich assay, suggesting that higher levels of multiplexing are possible. We used ProteinSeq to analyze proteins in plasma samples from cardiovascular disease (CVD) patient cohorts and matched controls. Three proteins, namely P-selectin, Cystatin-B and Kallikrein-6, were identified as putative diagnostic biomarkers for CVD. The latter two have not been previously reported in the literature and their potential roles must be validated in larger patient cohorts. We conclude that ProteinSeq is promising for screening large numbers of proteins and samples while the technology can provide a much-needed platform for validation of diagnostic markers in biobank samples and in clinical use. |
format | Online Article Text |
id | pubmed-3183061 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-31830612011-10-06 ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing Darmanis, Spyros Nong, Rachel Yuan Vänelid, Johan Siegbahn, Agneta Ericsson, Olle Fredriksson, Simon Bäcklin, Christofer Gut, Marta Heath, Simon Gut, Ivo Glynne Wallentin, Lars Gustafsson, Mats G. Kamali-Moghaddam, Masood Landegren, Ulf PLoS One Research Article Despite intense interest, methods that provide enhanced sensitivity and specificity in parallel measurements of candidate protein biomarkers in numerous samples have been lacking. We present herein a multiplex proximity ligation assay with readout via realtime PCR or DNA sequencing (ProteinSeq). We demonstrate improved sensitivity over conventional sandwich assays for simultaneous analysis of sets of 35 proteins in 5 µl of blood plasma. Importantly, we observe a minimal tendency to increased background with multiplexing, compared to a sandwich assay, suggesting that higher levels of multiplexing are possible. We used ProteinSeq to analyze proteins in plasma samples from cardiovascular disease (CVD) patient cohorts and matched controls. Three proteins, namely P-selectin, Cystatin-B and Kallikrein-6, were identified as putative diagnostic biomarkers for CVD. The latter two have not been previously reported in the literature and their potential roles must be validated in larger patient cohorts. We conclude that ProteinSeq is promising for screening large numbers of proteins and samples while the technology can provide a much-needed platform for validation of diagnostic markers in biobank samples and in clinical use. Public Library of Science 2011-09-29 /pmc/articles/PMC3183061/ /pubmed/21980495 http://dx.doi.org/10.1371/journal.pone.0025583 Text en Darmanis et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Darmanis, Spyros Nong, Rachel Yuan Vänelid, Johan Siegbahn, Agneta Ericsson, Olle Fredriksson, Simon Bäcklin, Christofer Gut, Marta Heath, Simon Gut, Ivo Glynne Wallentin, Lars Gustafsson, Mats G. Kamali-Moghaddam, Masood Landegren, Ulf ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing |
title | ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing |
title_full | ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing |
title_fullStr | ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing |
title_full_unstemmed | ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing |
title_short | ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing |
title_sort | proteinseq: high-performance proteomic analyses by proximity ligation and next generation sequencing |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3183061/ https://www.ncbi.nlm.nih.gov/pubmed/21980495 http://dx.doi.org/10.1371/journal.pone.0025583 |
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