Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability. / Assarsson, Erika; Lundberg, Martin; Holmquist, Göran; Björkesten, Johan; Thorsen, Stine Buch; Ekman, Daniel; Eriksson, Anna; Rennel Dickens, Emma; Ohlsson, Sandra; Edfeldt, Gabriella; Andersson, Ann-Catrin; Lindstedt, Patrik; Stenvang, Jan; Gullberg, Mats; Fredriksson, Simon.

In: PLOS ONE, Vol. 9, No. 4, e95192, 2014.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Assarsson, E, Lundberg, M, Holmquist, G, Björkesten, J, Thorsen, SB, Ekman, D, Eriksson, A, Rennel Dickens, E, Ohlsson, S, Edfeldt, G, Andersson, A-C, Lindstedt, P, Stenvang, J, Gullberg, M & Fredriksson, S 2014, 'Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability', PLOS ONE, vol. 9, no. 4, e95192. https://doi.org/10.1371/journal.pone.0095192

APA

Assarsson, E., Lundberg, M., Holmquist, G., Björkesten, J., Thorsen, S. B., Ekman, D., ... Fredriksson, S. (2014). Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability. PLOS ONE, 9(4), [e95192]. https://doi.org/10.1371/journal.pone.0095192

Vancouver

Assarsson E, Lundberg M, Holmquist G, Björkesten J, Thorsen SB, Ekman D et al. Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability. PLOS ONE. 2014;9(4). e95192. https://doi.org/10.1371/journal.pone.0095192

Author

Assarsson, Erika ; Lundberg, Martin ; Holmquist, Göran ; Björkesten, Johan ; Thorsen, Stine Buch ; Ekman, Daniel ; Eriksson, Anna ; Rennel Dickens, Emma ; Ohlsson, Sandra ; Edfeldt, Gabriella ; Andersson, Ann-Catrin ; Lindstedt, Patrik ; Stenvang, Jan ; Gullberg, Mats ; Fredriksson, Simon. / Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability. In: PLOS ONE. 2014 ; Vol. 9, No. 4.

Bibtex

@article{de4a45be09fc4cba98daef7fcabcb4ff,
title = "Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability",
abstract = "Medical research is developing an ever greater need for comprehensive high-quality data generation to realize the promises of personalized health care based on molecular biomarkers. The nucleic acid proximity-based methods proximity ligation and proximity extension assays have, with their dual reporters, shown potential to relieve the shortcomings of antibodies and their inherent cross-reactivity in multiplex protein quantification applications. The aim of the present study was to develop a robust 96-plex immunoassay based on the proximity extension assay (PEA) for improved high throughput detection of protein biomarkers. This was enabled by: (1) a modified design leading to a reduced number of pipetting steps compared to the existing PEA protocol, as well as improved intra-assay precision; (2) a new enzymatic system that uses a hyper-thermostabile enzyme, Pwo, for uniting the two probes allowing for room temperature addition of all reagents and improved the sensitivity; (3) introduction of an inter-plate control and a new normalization procedure leading to improved inter-assay precision (reproducibility). The multiplex proximity extension assay was found to perform well in complex samples, such as serum and plasma, and also in xenografted mice and resuspended dried blood spots, consuming only 1 µL sample per test. All-in-all, the development of the current multiplex technique is a step toward robust high throughput protein marker discovery and research.",
author = "Erika Assarsson and Martin Lundberg and G{\"o}ran Holmquist and Johan Bj{\"o}rkesten and Thorsen, {Stine Buch} and Daniel Ekman and Anna Eriksson and {Rennel Dickens}, Emma and Sandra Ohlsson and Gabriella Edfeldt and Ann-Catrin Andersson and Patrik Lindstedt and Jan Stenvang and Mats Gullberg and Simon Fredriksson",
note = "OA",
year = "2014",
doi = "10.1371/journal.pone.0095192",
language = "English",
volume = "9",
journal = "P L o S One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "4",

}

RIS

TY - JOUR

T1 - Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability

AU - Assarsson, Erika

AU - Lundberg, Martin

AU - Holmquist, Göran

AU - Björkesten, Johan

AU - Thorsen, Stine Buch

AU - Ekman, Daniel

AU - Eriksson, Anna

AU - Rennel Dickens, Emma

AU - Ohlsson, Sandra

AU - Edfeldt, Gabriella

AU - Andersson, Ann-Catrin

AU - Lindstedt, Patrik

AU - Stenvang, Jan

AU - Gullberg, Mats

AU - Fredriksson, Simon

N1 - OA

PY - 2014

Y1 - 2014

N2 - Medical research is developing an ever greater need for comprehensive high-quality data generation to realize the promises of personalized health care based on molecular biomarkers. The nucleic acid proximity-based methods proximity ligation and proximity extension assays have, with their dual reporters, shown potential to relieve the shortcomings of antibodies and their inherent cross-reactivity in multiplex protein quantification applications. The aim of the present study was to develop a robust 96-plex immunoassay based on the proximity extension assay (PEA) for improved high throughput detection of protein biomarkers. This was enabled by: (1) a modified design leading to a reduced number of pipetting steps compared to the existing PEA protocol, as well as improved intra-assay precision; (2) a new enzymatic system that uses a hyper-thermostabile enzyme, Pwo, for uniting the two probes allowing for room temperature addition of all reagents and improved the sensitivity; (3) introduction of an inter-plate control and a new normalization procedure leading to improved inter-assay precision (reproducibility). The multiplex proximity extension assay was found to perform well in complex samples, such as serum and plasma, and also in xenografted mice and resuspended dried blood spots, consuming only 1 µL sample per test. All-in-all, the development of the current multiplex technique is a step toward robust high throughput protein marker discovery and research.

AB - Medical research is developing an ever greater need for comprehensive high-quality data generation to realize the promises of personalized health care based on molecular biomarkers. The nucleic acid proximity-based methods proximity ligation and proximity extension assays have, with their dual reporters, shown potential to relieve the shortcomings of antibodies and their inherent cross-reactivity in multiplex protein quantification applications. The aim of the present study was to develop a robust 96-plex immunoassay based on the proximity extension assay (PEA) for improved high throughput detection of protein biomarkers. This was enabled by: (1) a modified design leading to a reduced number of pipetting steps compared to the existing PEA protocol, as well as improved intra-assay precision; (2) a new enzymatic system that uses a hyper-thermostabile enzyme, Pwo, for uniting the two probes allowing for room temperature addition of all reagents and improved the sensitivity; (3) introduction of an inter-plate control and a new normalization procedure leading to improved inter-assay precision (reproducibility). The multiplex proximity extension assay was found to perform well in complex samples, such as serum and plasma, and also in xenografted mice and resuspended dried blood spots, consuming only 1 µL sample per test. All-in-all, the development of the current multiplex technique is a step toward robust high throughput protein marker discovery and research.

U2 - 10.1371/journal.pone.0095192

DO - 10.1371/journal.pone.0095192

M3 - Journal article

VL - 9

JO - P L o S One

JF - P L o S One

SN - 1932-6203

IS - 4

M1 - e95192

ER -

ID: 125944652