Technical performance of a proximity extension assay inflammation biomarker panel with synovial fluid
Struglics, André LU ; Larsson, Staffan LU
; Lohmander, L Stefan
LU
and Swärd, Per
LU
(2022)
In Osteoarthritis and Cartilage Open
4(3).
- Abstract
- The plasma proteome can provide a deeper understanding of the pathophysiology of different diseases and is a key source for detection of new biomarkers. Proximity extension assay (PEA), developed by Olink Proteomics (Uppsala, Sweden), detects proteins by using pairs of antibodies that are linked to oligonucleotides; upon target binding, the probes anneal when in proximity and the oligonucleotides are extended by DNA polymerase and the newly formed antigen is quantified by real-time PCR [1]. PEA provides high specificity and sensitivity, and the possibility to measure the relative abundance of a large number of proteins (92 up to 384 biomarkers), only using a few μL of biofluid per sample. This technique makes it possible to assess... (More)
- The plasma proteome can provide a deeper understanding of the pathophysiology of different diseases and is a key source for detection of new biomarkers. Proximity extension assay (PEA), developed by Olink Proteomics (Uppsala, Sweden), detects proteins by using pairs of antibodies that are linked to oligonucleotides; upon target binding, the probes anneal when in proximity and the oligonucleotides are extended by DNA polymerase and the newly formed antigen is quantified by real-time PCR [1]. PEA provides high specificity and sensitivity, and the possibility to measure the relative abundance of a large number of proteins (92 up to 384 biomarkers), only using a few μL of biofluid per sample. This technique makes it possible to assess low-abundant proteins which are not accessible using mass spectrometry techniques [2]. The multi-assay/plex capacity of PEA is comparable with other multiplex assays like Meso Scale Discovery (MSD); MSD uses electrochemiluminescence measuring the concentration of up to 10 biomarkers per well in 25 μl per sample. The PEA has an advantage over enzyme-linked immunosorbent assay (ELISA) and MSD assays in that PEA has a high number of biomarkers which are quantifiable in very low sample volume.
In plasma and serum, the correlation between the PEA technique and other techniques, including ELISA and electrochemiluminescence has been shown to be strong [[3], [4], [5], [6], [7], [8]]. The technical performance of the PEA technique in synovial fluid is unknown. The characteristics of synovial fluid differs from plasma in that some proteins are more abundant, whereas other proteins are less abundant. Compared to plasma, synovial fluid is very viscous due to high levels of hyaluronic acid [9]. These differences could interfere with the PEA analysis technique. To interpret protein data generated by the PEA technique in synovial fluid, information on the technical performance of this assay on synovial fluid samples is needed [10].
The purpose of the present study was to evaluate the technical performance of Olink's inflammation PEA with synovial fluid samples and compare it with the performance of the same PEA in the more commonly used biofluids, serum and plasma. The second aim was to compare some of the PEA data with data obtained by immunoassays using MSD. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/2323620e-f3ea-4f41-8f93-935756036299
- author
- Struglics, André
LU
; Larsson, Staffan
LU
; Lohmander, L Stefan
LU
and Swärd, Per
LU
- organization
- publishing date
- 2022-09
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Osteoarthritis and Cartilage Open
- volume
- 4
- issue
- 3
- article number
- 100293
- publisher
- Elsevier
- external identifiers
-
- scopus:85135518985
- pmid:36474941
- ISSN
- 2665-9131
- DOI
- 10.1016/j.ocarto.2022.100293
- language
- English
- LU publication?
- yes
- id
- 2323620e-f3ea-4f41-8f93-935756036299
- date added to LUP
- 2023-04-14 12:20:24
- date last changed
- 2024-04-19 20:45:13
@article{2323620e-f3ea-4f41-8f93-935756036299,
abstract = {{The plasma proteome can provide a deeper understanding of the pathophysiology of different diseases and is a key source for detection of new biomarkers. Proximity extension assay (PEA), developed by Olink Proteomics (Uppsala, Sweden), detects proteins by using pairs of antibodies that are linked to oligonucleotides; upon target binding, the probes anneal when in proximity and the oligonucleotides are extended by DNA polymerase and the newly formed antigen is quantified by real-time PCR [1]. PEA provides high specificity and sensitivity, and the possibility to measure the relative abundance of a large number of proteins (92 up to 384 biomarkers), only using a few μL of biofluid per sample. This technique makes it possible to assess low-abundant proteins which are not accessible using mass spectrometry techniques [2]. The multi-assay/plex capacity of PEA is comparable with other multiplex assays like Meso Scale Discovery (MSD); MSD uses electrochemiluminescence measuring the concentration of up to 10 biomarkers per well in 25 μl per sample. The PEA has an advantage over enzyme-linked immunosorbent assay (ELISA) and MSD assays in that PEA has a high number of biomarkers which are quantifiable in very low sample volume.<br/><br/>In plasma and serum, the correlation between the PEA technique and other techniques, including ELISA and electrochemiluminescence has been shown to be strong [[3], [4], [5], [6], [7], [8]]. The technical performance of the PEA technique in synovial fluid is unknown. The characteristics of synovial fluid differs from plasma in that some proteins are more abundant, whereas other proteins are less abundant. Compared to plasma, synovial fluid is very viscous due to high levels of hyaluronic acid [9]. These differences could interfere with the PEA analysis technique. To interpret protein data generated by the PEA technique in synovial fluid, information on the technical performance of this assay on synovial fluid samples is needed [10].<br/><br/>The purpose of the present study was to evaluate the technical performance of Olink's inflammation PEA with synovial fluid samples and compare it with the performance of the same PEA in the more commonly used biofluids, serum and plasma. The second aim was to compare some of the PEA data with data obtained by immunoassays using MSD.}},
author = {{Struglics, André and Larsson, Staffan and Lohmander, L Stefan and Swärd, Per}},
issn = {{2665-9131}},
language = {{eng}},
number = {{3}},
publisher = {{Elsevier}},
series = {{Osteoarthritis and Cartilage Open}},
title = {{Technical performance of a proximity extension assay inflammation biomarker panel with synovial fluid}},
url = {{http://dx.doi.org/10.1016/j.ocarto.2022.100293}},
doi = {{10.1016/j.ocarto.2022.100293}},
volume = {{4}},
year = {{2022}},
}