Development and Application of Proximity Assays for Proteome Analysis in Medicine

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MARC

de Oliveira, Felipe Marques SouzaUppsala universitet,Molekylära verktyg (author)

Development and Application of Proximity Assays for Proteome Analysis in Medicine

BookEnglish2018

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Uppsala :Acta Universitatis Upsaliensis,2018
60 s.
electronicrdacarrier

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LIBRIS-ID:oai:DiVA.org:uu-334536
ISBN:9789151301648
https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-334536URI

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Language:English
Summary in:English

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Subject category:vet swepub-contenttype
Subject category:dok swepub-publicationtype

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Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine,1651-6206 ;1400

Notes

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.

Subject headings and genre

MEDICIN OCH HÄLSOVETENSKAP Medicinsk bioteknologi Medicinsk bioteknologi hsv//swe
MEDICAL AND HEALTH SCIENCES Medical Biotechnology Medical Biotechnology hsv//eng
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

Added entries (persons, corporate bodies, meetings, titles ...)

Kamali-Moghaddam, MasoodUppsala universitet,Molekylära verktyg (thesis advisor)
Landegren, UlfUppsala universitet,Molekylära verktyg (thesis advisor)
Gullberg, MatsQ-Linea (thesis advisor)
Lehtiö, Janne,ProfessorKarolinska Universitet, Department of Oncology-Pathology (OnkPat), K7 (opponent)
Uppsala universitetMolekylära verktyg (creator_code:org_t)

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