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Detection of post-translational modification of cancer biomarkers via proximity ligation assay
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology.
Univ Porto, i3S, Oporto, Portugal.;Univ Porto, IPATIMUP Inst Mol Pathol & Immunol, Oporto, Portugal..
Univ Copenhagen, Dept Chem, Copenhagen, Denmark..
Univ Copenhagen, Dept Chem, Copenhagen, Denmark..
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2016 (English)In: Glycobiology, ISSN 0959-6658, E-ISSN 1460-2423, Vol. 26, no 12, 1455-1455 p.Article in journal, Meeting abstract (Refereed) Published
Place, publisher, year, edition, pages
2016. Vol. 26, no 12, 1455-1455 p.
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:uu:diva-316637ISI: 000392935600200OAI: oai:DiVA.org:uu-316637DiVA: diva2:1078638
Conference
Annual Meeting of the Society-for-Glycobiology, NOV 19-22, 2016, New Orleans, LA
Available from: 2017-03-06 Created: 2017-03-06 Last updated: 2017-11-29Bibliographically approved
In thesis
1. Development and Application of Proximity Assays for Proteome Analysis in Medicine
Open this publication in new window or tab >>Development and Application of Proximity Assays for Proteome Analysis in Medicine
2018 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Along with proteins, a myriad of different molecular biomarkers, such as post-translational modifications and autoantibodies, could be used in an attempt to improve disease detection and progression. In this thesis, I build on several iterations of the proximity ligation assay to develop and apply new adaptable methods to facilitate detection of proteins, autoantibodies and post-translational modifications.

In paper I, we present an adaptation of the solid-phase proximity ligation assay (SP-PLA) for the detection of post-translational modification of proteins (PTMs). The assay was adapted for the detection of two of the most commons PTMs present in proteins, glycosylation and phosphorylation, offering the encouraging prospect of using detection of PTMs in a diagnostic or prognostic capacity. 

In paper II, we developed a variant of the proximity ligation assay using micro titer plate for detection and quantification of protein using optical density as readout in the fluorometer, termed PLARCA. With a detection limit considerably lower than ELISA, PLARCA detected femtomolar levels of these proteins in patient samples.

In paper III, we aim to compare detection values of samples collected from earlobe capillary, venous plasma, as well as capillary plasma stored in dried plasma spots (DPS) assessed with a 92-plex inflammation panel using multiplex proximity extension assay (PEA). Despite the high variability in protein measurements between the three sample sources, we were able to conclude that earlobe capillary sampling is a suitable less invasive alternative, to venipuncture.

In paper IV, we describe the application of PLARCA and proximity extension assay (PEA) for the detection of GAD65 autoantibodies (GADA). Thus, offering highly sensitive and specific autoimmunity detection.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2018. 60 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 1400
Keyword
Solid-phase proximity ligation assay, post-translational modifications, glycosylation, phosphorylation, Enzyme-linked immunosorbent assay, immunoassay and rolling circle amplification, Proximity Extension Assay; inflammation protein biomarkers, autoantibodies; autoimmune disease
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:uu:diva-334536 (URN)978-91-513-0164-8 (ISBN)
Public defence
2018-01-18, B:41, BMC, Husargatan 3, Uppsala, 09:00 (English)
Opponent
Supervisors
Available from: 2017-12-15 Created: 2017-11-23 Last updated: 2017-12-15

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de Oliveira, Felipe Marques SouzaKamali-Moghaddam, Masood

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