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Systematic assessme...
Systematic assessment of antibody selectivity in plasma based on a resource of enrichment profiles
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- Fredolini, Claudia (author)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab,KTH Royal Inst Technol, Dept Prot Sci, Div Affin Prote, Sci Life Lab, S-17121 Solna, Sweden
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- Byström, Sanna (author)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab,KTH Royal Inst Technol, Dept Prot Sci, Div Affin Prote, Sci Life Lab, S-17121 Solna, Sweden
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- Sanchez-Rivera, Laura (author)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab,KTH Royal Inst Technol, Dept Prot Sci, Div Affin Prote, Sci Life Lab, S-17121 Solna, Sweden
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- Ioannou, Marina (author)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab,KTH Royal Inst Technol, Dept Prot Sci, Div Affin Prote, Sci Life Lab, S-17121 Solna, Sweden
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- Tamburro, Davide (author)
- Karolinska Inst, Sci Life Lab, Dept Oncol Pathol, Canc Prote, S-17121 Solna, Sweden.
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- Pontén, Fredrik (author)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Klinisk och experimentell patologi
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- Branca, Rui M. (author)
- Karolinska Institutet
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- Nilsson, Peter (author)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab,KTH Royal Inst Technol, Dept Prot Sci, Div Affin Prote, Sci Life Lab, S-17121 Solna, Sweden
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- Lehtio, Janne (author)
- Karolinska Institutet
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- Schwenk, Jochen M. (author)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab,KTH Royal Inst Technol, Dept Prot Sci, Div Affin Prote, Sci Life Lab, S-17121 Solna, Sweden
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(creator_code:org_t)
- 2019-06-06
- 2019
- English.
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In: Scientific Reports. - : Nature Publishing Group. - 2045-2322. ; 9
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https://doi.org/10.1...
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https://uu.diva-port... (primary) (Raw object)
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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http://kipublication...
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Abstract
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- There is a strong need for procedures that enable context and application dependent validation of antibodies. Here, we applied a magnetic bead assisted workflow and immunoprecipitation mass spectrometry (IP-MS/MS) to assess antibody selectivity for the detection of proteins in human plasma. A resource was built on 414 IP experiments using 157 antibodies (targeting 120 unique proteins) in assays with heat-treated or untreated EDTA plasma. For each protein we determined their antibody related degrees of enrichment using z-scores and their frequencies of identification across all IP assays. Out of 1,313 unique endogenous proteins, 426 proteins (33%) were detected in >20% of IPs, and these background components were mainly comprised of proteins from the complement system. For 45% (70/157) of the tested antibodies, the expected target proteins were enriched (z-score >= 3). Among these 70 antibodies, 59 (84%) co-enriched other proteins beside the intended target and mainly due to sequence homology or protein abundance. We also detected protein interactions in plasma, and for IGFBP2 confirmed these using several antibodies and sandwich immunoassays. The protein enrichment data with plasma provide a very useful and yet lacking resource for the assessment of antibody selectivity. Our insights will contribute to a more informed use of affinity reagents for plasma proteomics assays.
Subject headings
- NATURVETENSKAP -- Biologi -- Bioinformatik och systembiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Bioinformatics and Systems Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Immunologi inom det medicinska området (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Immunology in the medical area (hsv//eng)
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- By the author/editor
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Fredolini, Claud ...
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Byström, Sanna
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Sanchez-Rivera, ...
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Ioannou, Marina
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Tamburro, Davide
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Pontén, Fredrik
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show more...
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Branca, Rui M.
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Nilsson, Peter
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Lehtio, Janne
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Schwenk, Jochen ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Bioinformatics a ...
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Medical Biotechn ...
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and Medical Biotechn ...
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Basic Medicine
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and Immunology in th ...
- Articles in the publication
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Scientific Repor ...
- By the university
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Royal Institute of Technology
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Uppsala University
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Karolinska Institutet