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Antibody-based bead arrays for high-throughput protein profiling in human plasma and serum

Drobin, Kimi (author)
KTH,Proteinvetenskap
Schwenk, Jochen M. (thesis advisor)
KTH,Science for Life Laboratory, SciLifeLab,Proteinvetenskap
Nilsson, Peter (thesis advisor)
KTH,Science for Life Laboratory, SciLifeLab
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Qundos, Ulrika (thesis advisor)
KTH,Science for Life Laboratory, SciLifeLab
Silveira, Angela (opponent)
Pernemalm, Maria (opponent)
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 (creator_code:org_t)
ISBN 9789177297390
KTH Royal Institute of Technology, 2018
English 58 s.
Series: TRITA-CBH-FOU ; 2018:9
  • Licentiate thesis (other academic/artistic)
Abstract Subject headings
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  • Affinity-based proteomics utilizes affinity binders to detect target proteins in a large-scale manner. This thesis describes a high-throughput method, which enables the search for biomarker candidates in human plasma and serum. A highly multiplexed antibody-based suspension bead array is created by coupling antibodies generated in the Human Protein Atlas project to color-coded beads. The beads are combined for parallel analysis of up to 384 analytes in patient and control samples. This provides data to compare protein levels from the different groups.In paper I osteoporosis patients are compared to healthy individuals to find disease-linked proteins. An untargeted discovery screening was conducted using 4608 antibodies in 16 cases and 6 controls. This revealed 72 unique proteins, which appeared differentially abundant. A validation screening of 91 cases and 89 controls confirmed that the protein autocrine motility factor receptor (AMFR) is decreased in the osteoporosis patients.Paper II investigates the risk proteome of inflammatory bowel disease (IBD). Antibodies targeting 209 proteins corresponding to 163 IBD genetic risk loci were selected. To find proteins related to IBD or its subgroups, sera from 49 patients with Crohn’s disease, 51 with ulcerative colitis and 50 matched controls were analyzed. From these targeted assays, the known inflammation-related marker serum amyloid protein A (SAA) was shown to be elevated in the IBD cases. In addition, the protein laccase (multi-copper oxidoreductase) domain containing 1 (LACC1) was found to be decreased in the IBD subjects.In conclusion, assays using affinity-based bead arrays were developed and applied to screen human plasma and serum samples in two disease contexts. Untargeted and targeted screening strategies were applied to discover disease-associated proteins. Upon further validation, these potential biomarker candidates could be valuable in future disease studies.

Subject headings

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

Keyword

proteomics
affinity proteomics
immunoassay
antibody microarray
suspension bead array
protein profiling
plasma
serum
biomarker discovery
osteoporosis
inflammatory bowel disease
Crohn's disease
ulcerative colitis
Medical Technology
Medicinsk teknologi

Publication and Content Type

vet (subject category)
lic (subject category)

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Drobin, Kimi
Schwenk, Jochen ...
Nilsson, Peter
Qundos, Ulrika
Silveira, Angela
Pernemalm, Maria
About the subject
MEDICAL AND HEALTH SCIENCES
MEDICAL AND HEAL ...
and Medical Biotechn ...
and Medical Biotechn ...
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