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Development and App...
Development and Application of Proximity Assays for Proteome Analysis in Medicine
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- de Oliveira, Felipe Marques Souza (författare)
- Uppsala universitet,Molekylära verktyg
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- Kamali-Moghaddam, Masood (preses)
- Uppsala universitet,Molekylära verktyg
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- Landegren, Ulf (preses)
- Uppsala universitet,Molekylära verktyg
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- Gullberg, Mats (preses)
- Q-Linea
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- Lehtiö, Janne, Professor (opponent)
- Karolinska Universitet, Department of Oncology-Pathology (OnkPat), K7
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(creator_code:org_t)
- ISBN 9789151301648
- Uppsala : Acta Universitatis Upsaliensis, 2018
- Engelska 60 s.
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Serie: Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, 1651-6206 ; 1400
- Relaterad länk:
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Abstract
Ämnesord
Stäng
- 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.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Nyckelord
- 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
Publikations- och innehållstyp
- vet (ämneskategori)
- dok (ämneskategori)
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