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Proximity extension assay PEA

Megan E Roerink, Hans Knoop, Ewald M Bronkhorst, Henk A Mouthaan, Luuk J A C Hawinkels, Leo A B Joosten, Jos W M van der Meer
BACKGROUND: Cytokine disturbances have been suggested to be associated with the Chronic Fatigue Syndrome/Myalgic encephalomyelitis (CFS/ME) for decades. METHODS: Fifty female CFS patients were included in a study on the effect of the interleukin-1-receptor antagonist anakinra or placebo during 4 weeks. EDTA plasma was collected from patients before and directly after treatment. At baseline, plasma samples were collected at the same time from 48 healthy, age-matched female neighborhood controls...
December 29, 2017: Journal of Translational Medicine
Jeneen Panezai, Ambereen Ghaffar, Mohammad Altamash, Karl-Gösta Sundqvist, Per-Erik Engström, Anders Larsson
BACKGROUND: Periodontal disease (PD) is characterized by inflammatory tissue destruction in tooth supporting apparatus. Many studies indicate that the underlying pathogenesis is in concordance with rheumatoid arthritis (RA) sharing immune-inflammatory events affect both diseases. The aim of this study was to investigate serum cytokines, chemokines, growth factors, enzymes and costimulatory proteins in association with periodontal conditions in PD and RA subjects. MATERIALS & METHODS: Periodontal examination was performed in RA (n = 38), PD (n = 38) and healthy subjects (n = 14)...
2017: PloS One
Erik Andersson, Daniel Bergemalm, Robert Kruse, Gunter Neumann, Mauro D'Amato, Dirk Repsilber, Jonas Halfvarson
OBJECTIVE: Genetic and immunological data indicate that inflammatory bowel disease (IBD) are characterized by specific inflammatory protein profiles. However, the serum proteome of IBD is still to be defined. We aimed to characterize the inflammatory serum protein profiles of Crohn's disease (CD) and ulcerative colitis (UC), using the novel proximity extension assay. METHODS: A panel of 91 inflammatory proteins were quantified in a discovery cohort of CD (n = 54), UC patients (n = 54), and healthy controls (HCs; n = 54)...
2017: PloS One
Johan Björkesten, Stefan Enroth, Qiujin Shen, Lotta Wik, David M Hougaard, Arieh S Cohen, Lene Sörensen, Vilmantas Giedraitis, Martin Ingelsson, Anders Larsson, Masood Kamali-Moghaddam, Ulf Landegren
An important motivation for the construction of biobanks is to discover biomarkers that identify diseases at early, potentially curable stages. This will require biobanks from large numbers of individuals, preferably sampled repeatedly, where the samples are collected and stored under conditions that preserve potential biomarkers. Dried blood samples are attractive for biobanking because of the ease and low cost of collection and storage. Here we have investigated their suitability for protein measurements...
July 2017: Molecular & Cellular Proteomics: MCP
Björn Gerdle, Bijar Ghafouri, Nazdar Ghafouri, Emmanuel Bäckryd, Torsten Gordh
This cross-sectional study investigates the plasma inflammatory profile of chronic widespread pain (CWP) patients compared to healthy controls (CON). Rather than analyzing a relatively few substances at a time, we used a new multiplex proximity extension assay (PEA) panel that enabled the simultaneous analysis of 92 inflammation-related proteins, mainly cytokines and chemokines.Seventeen women with CWP and 21 female CON participated and a venous blood sample was drawn from all subjects. Pain intensity and pain thresholds for pressure, heat, and cold were registered...
March 2017: Medicine (Baltimore)
Pia Larssen, Lotta Wik, Paulo Czarnewski, Maria Eldh, Liza Löf, K Göran Ronquist, Louise Dubois, Eva Freyhult, Caroline J Gallant, Johan Oelrich, Anders Larsson, Gunnar Ronquist, Eduardo J Villablanca, Ulf Landegren, Susanne Gabrielsson, Masood Kamali-Moghaddam
Extracellular vesicles (EVs) are membrane-coated objects such as exosomes and microvesicles, released by many cell-types. Their presence in body fluids and the variable surface composition and content render them attractive potential biomarkers. The ability to determine their cellular origin could greatly move the field forward. We used multiplex proximity extension assays (PEA) to identify with high specificity and sensitivity the protein profiles of exosomes of different origins, including seven cell lines and two different body fluids...
March 2017: Molecular & Cellular Proteomics: MCP
Katarina Edfeldt, Kosmas Daskalakis, Christofer Bäcklin, Olov Norlén, Eva Tiensuu Janson, Gunnar Westin, Per Hellman, Peter Stålberg
Small intestinal neuroendocrine tumors (SI-NETs) are amine- and peptide-producing neoplasms. Most patients display metastases at the time of diagnosis; they have an unpredictable individual disease course and the tumors are often therapy resistant. Chromogranin A and 5-hydroxyindoleacetic acid are the biomarkers clinically used most often today, but there is a great need for novel diagnostic and prognostic biomarkers and new therapeutic targets. Sixty-nine biomarkers were screened in serum from 23 SI-NET patients and 23 healthy controls using the multiplex proximity ligation assay (PLA)...
2017: Neuroendocrinology
Lars Lind, Johan Sundström, Markus Stenemo, Emil Hagström, Johan Ärnlöv
BACKGROUND: Apart from several established clinical risk factors for atrial fibrillation (AF), a number of biomarkers have also been identified as potential risk factors for AF. None of these have so far been adopted in clinical practice. OBJECTIVE: To use a novel custom-made proteomics chip to discover new prognostic biomarkers for AF risk. METHODS: In two independent community-based cohorts (Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) study (978 participants without AF, mean age 70...
March 2017: Heart: Official Journal of the British Cardiac Society
Caitríona Grönberg, Giuseppe Asciutto, Ana Persson, Gunilla Nordin Fredrikson, Jan Nilsson, Isabel Gonçalves, Harry Björkbacka
BACKGROUND AND PURPOSE: Increased interleukin 16 (IL-16) levels in carotid plaques have been associated with reduced incidence of cardiovascular (CV) events during follow-up in patients who underwent carotid endarterectomy (CEA). In the present study we aimed to determine whether high circulating levels of IL-16 also are associated with a decreased risk of CV events after CEA. METHODS: Patients, who had their carotid plaques surgically removed (n=473), were followed for a mean follow-up time of 3...
September 2016: Cytokine
Aurora Moen, Anne-Li Lind, Måns Thulin, Masood Kamali-Moghaddam, Cecilie Røe, Johannes Gjerstad, Torsten Gordh
Earlier studies suggest that lumbar radicular pain following disc herniation may be associated with a local or systemic inflammatory process. In the present study, we investigated the serum inflammatory protein profile of such patients. All 45 patients were recruited from Oslo University Hospital, Ullevål, Norway, during the period 2007-2009. The new multiplex proximity extension assay (PEA) technology was used to analyze the levels of 92 proteins. Interestingly, the present data showed that patients with radicular pain 12 months after disc herniation may be different from other patients with regard to many measurable serum cytokines...
2016: International Journal of Inflammation
Anders Larsson, Lena Carlsson, Anne-Li Lind, Torsten Gordh, Constantin Bodolea, Masood Kamali-Moghaddam, Måns Thulin
Cytokines and chemokines regulate many functions in the body including the brain. The interactions between adipose tissue and the central nervous system (CNS) are important for the regulation of energy balance. CNS function is also influenced by age. The aim of the present study was to investigate the effects of body mass index (BMI) and age on cytokine and chemokine levels in cerebrospinal fluid. Cerebrospinal fluid samples (n=89) were collected from patients undergoing routine surgical procedures. The samples were analyzed using the multiplex proximity extension assay (PEA) in which 92 different cytokines are measured simultaneously using minute sample volume...
December 2015: Cytokine
Anders Larsson, Lena Carlsson, Torsten Gordh, Anne-Li Lind, Måns Thulin, Masood Kamali-Moghaddam
Cytokines play important roles as regulators of cell functions, and over the last decades a number of cytokine assays have been developed. The aim of the present study was to investigate the effects of age and gender on a large number of cytokines. Plasma samples were collected from 33 healthy blood donors. The samples were analyzed using a multiplex proximity extension assay (PEA) allowing simultaneous measurement of 92 cytokines and four technical controls. Biomarkers with less than 80% quantitative results were excluded leaving 63 cytokines that were analyzed for the effects of gender and age...
October 2015: Journal of Immunological Methods
Erika Assarsson, Martin Lundberg, Göran Holmquist, Johan Björkesten, Stine Bucht Thorsen, Daniel Ekman, Anna Eriksson, Emma Rennel Dickens, Sandra Ohlsson, Gabriella Edfeldt, Ann-Catrin Andersson, Patrik Lindstedt, Jan Stenvang, Mats Gullberg, Simon Fredriksson
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...
2014: PloS One
Feng Gao, Suna P Gulay, Wojciech Kasprzak, Jonathan D Dinman, Bruce A Shapiro, Anne E Simon
The Pea enation mosaic virus (PEMV) 3' translational enhancer, known as the kissing-loop T-shaped structure (kl-TSS), binds to 40S subunits, 60S subunits, and 80S ribosomes, whereas the Turnip crinkle virus (TCV) TSS binds only to 60S subunits and 80S ribosomes. Using electrophoretic mobility gel shift assay (EMSA)-based competition assays, the kl-TSS was found to occupy a different site in the ribosome than the P-site-binding TCV TSS, suggesting that these two TSS employ different mechanisms for enhancing translation...
November 2013: Journal of Virology
Liesbeth Van Wesenbeeck, Hanne Meeuws, Hans De Wolf, Lieven Stuyver
The rapid influenza diagnostic tests (RIDTs) are widely distributed, simple to use, but often lack sensitivity as compared to gold standard methods (viral culture and nucleic acid detection technologies). Applying RIDTs outside of epidemic or pandemic infections results in large numbers of false negatives. Hence, a sensitive RIDT that would reduce the number of false negatives would result in an increased clinical value. We evaluated the potential of a proximity extension assay (PEA) for the detection of influenza A H1 viruses...
October 2013: Journal of Virological Methods
Cassie Aldridge, Amanda Storm, Kenneth Cline, Carole Dabney-Smith
Twin arginine transport (Tat) systems transport folded proteins using proton-motive force as sole energy source. The thylakoid Tat system comprises three membrane components. A complex composed of cpTatC and Hcf106 is the twin arginine signal peptide receptor. Signal peptide binding triggers assembly of Tha4 for the translocation step. Tha4 is thought to serve as the protein-conducting element, and the topology it adopts during transport produces the transmembrane passageway. We analyzed Tha4 topology and conformation in actively transporting translocases and compared that with Tha4 in nontransporting membranes...
October 5, 2012: Journal of Biological Chemistry
Martin Lundberg, Anna Eriksson, Bonnie Tran, Erika Assarsson, Simon Fredriksson
Convenient and well-performing protein detection methods for a wide range of targets are in great demand for biomedical research and future diagnostics. Assays without the need for washing steps while still unaffected when analyzing complex biological samples are difficult to develop. Herein, we report a well-characterized nucleic acid proximity-based assay using antibodies, called Proximity Extension Assay (PEA), showing good performance in plasma samples. Target-specific antibody pairs are linked to DNA strands that upon simultaneous binding to the target analyte create a real-time PCR amplicon in a proximity-dependent manner enabled by the action of a DNA polymerase...
August 2011: Nucleic Acids Research
S A Hogenhout, F van der Wilk, M Verbeek, R W Goldbach, J F van den Heuvel
A GroEL homolog with a molecular mass of 60 kDa, produced by the primary endosymbiotic bacterium (a Buchnera sp.) of Myzus persicae and released into the hemolymph, has previously been shown to be a key protein in the transmission of potato leafroll virus (PLRV). Like other luteoviruses and pea enation mosaic virus, PLRV readily binds to extracellular Buchnera GroEL, and in vivo interference in this interaction coincides with reduced capsid integrity and loss of infectivity. To gain more knowledge of the nature of the association between PLRV and Buchnera GroEL, the groE operon of the primary endosymbiont of M...
January 1998: Journal of Virology
J Yee, G S Kuncio, B Bhandari, F S Shihab, E G Neilson
Stromelysin-1, matrix metalloproteinase-3 (MMP-3), is an important endopeptidase selectively expressed by somatic cells in organ tissues. The renal tubulointerstitium, for example, comprises tubular epithelium and interstitial fibroblasts forming the principal mass of the kidney. We observed that mRNA encoding stromelysin-1 is detectable in murine renal fibroblasts, but not in proximal tubular epithelium. Transcripts measured by RNase protection assay in renal fibroblasts increase following exposure to phorbol ester, and thereafter, activated stromelysin-1 protein can be detected in culture media by Western blotting...
July 1997: Kidney International
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