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Plasma Profiling Reveals Human Fibulin-1 as Candidate Marker for Renal Impairment
KTH, School of Biotechnology (BIO), Proteomics (closed 20130101). KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-1855-703X
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2011 (English)In: Journal of Proteome Research, ISSN 1535-3893, E-ISSN 1535-3907, Vol. 10, no 11, 4925-4934 p.Article in journal (Refereed) Published
Abstract [en]

There is a need for reliable and sensitive biomarkers for renal impairments to detect early signs of kidney toxicity and to monitor progression of disease. Here, antibody suspension bead arrays were applied to profile plasma samples from patients with four types of kidney disorders: glomerulonephritis, diabetic nephropathy, obstructive uropathy, and analgesic abuse. In total, 200 clinical renal-associated cases and control plasma samples from different cohorts were profiled. Parallel plasma protein profiles were obtained using biotinylated and nonfractionated samples and a selected set of 94 proteins targeted by 129 antigen-purified polyclonal antibodies. Out of the analyzed target proteins, human fibulin-1 was detected at significantly higher levels in the glomerulonephritis patient group compared to the controls and with elevated levels in patient samples for all other renal disorders investigated. Two polyclonal antibodies and one monoclonal antibody directed toward separate, nonoverlapping epitopes showed the same trend in the discovery cohorts. A technical verification using Western blot analysis of selected patient plasma confirmed the trends toward higher abundance of the target protein in disease samples. Furthermore, a verification study was carried out in the context of glomerulonephritis using an independent case and control cohort, and this confirmed the results from the discovery cohort, suggesting that plasma levels of fibulin-1 could serve as a potential indicator to monitor kidney malfunction or kidney damage.

Place, publisher, year, edition, pages
2011. Vol. 10, no 11, 4925-4934 p.
Keyword [en]
affinity proteomics, plasma profiling, antibody microarray, kidney disorders, biomarker, fibulin-1, glomerulonephritis
National Category
Biological Sciences
URN: urn:nbn:se:kth:diva-51288DOI: 10.1021/pr200286cISI: 000296414700004ScopusID: 2-s2.0-80655139676OAI: diva2:463894
Knut and Alice Wallenberg FoundationScience for Life Laboratory - a national resource center for high-throughput molecular bioscience
QC 20111212Available from: 2011-12-12 Created: 2011-12-12 Last updated: 2013-02-08Bibliographically approved
In thesis
1. Bead based protein profiling in blood
Open this publication in new window or tab >>Bead based protein profiling in blood
2013 (Swedish)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis is about protein profiling in blood-derived samples using suspension bead ar- rays built with protein affinity reagents, and the evaluation of binding characteristics and potential disease relation of such profiles.

A central aim of the presented work was to discover and verify disease associated protein profiles in blood-derived samples such as serum or plasma. This was based on immobiliz- ing antigens or antibodies on color-coded beads for a multiplexed analysis. This concept generally allow for a dual multiplexing because hundreds of samples can be screened for hundreds of proteins in a miniaturized and parallelized fashion. At first, protein antigens were used to study humoral immune responses in cattle suffering from a mycoplasma infec- tion (Paper I). Here, the most immunogenic of the applied antigens were identified based on reactivity profiles from the infected cattle, and were combined into an antigen cocktail to serve as a diagnostic assay in a standard ELISA set-up. Next, antibodies and their em- ployment in assays with directly labeled human samples was initiated. This procedure was applied in a study of kidney disorders where screening of plasma resulted in the discovery of a biomarker candidate, fibulin-1 (Paper II). In parallel to the disease related applica- tions, systematic evaluations of the protein profiles were conducted. Protein profiles from 2,300 antibodies were classified on the bases of binding properties in relation to sample heating and stringent washing (Paper III). With a particular focus on heat dependent de- tectability, a method was developed to visualize those proteins that were captured to the beads in an immunoassay by using Western blotting (Paper IV). In conclusion, this thesis presents examples of the possibilities of comparative plasma profiling enabled by protein bead arrays. 

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2013. 116 p.
Trita-BIO-Report, ISSN 1654-2312 ; 2013:4
Affinity proteomics, protein array, suspension bead array, antigen, antibody, biomarker discovery, serology, selectivity, sensitivity, serum, plasma
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
urn:nbn:se:kth:diva-117960 (URN)978-91-7501-629-0 (ISBN)
Public defence
2013-03-01, Gardaulan, Smittskyddsinstitutet, Nobels väg 18, Solna, 10:00 (English)

QC 20130208

Available from: 2013-02-08 Created: 2013-02-07 Last updated: 2013-02-08Bibliographically approved

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Neiman, MajaUhlén, MathiasSchwenk, Jochen M.Nilsson, Peter
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