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Advancing the immunoaffinity platform AFFIRM to targeted measurements of proteins in serum in the pg/ml range

Säll, Anna LU ; Corbee, Daniel; Vikström, Sara; Ottosson, Filip LU ; Persson, Helena and Waldemarson, Sofia LU (2018) In PLoS ONE 13(2).
Abstract

There is a great need for targeted protein assays with the capacity of sensitive measurements in complex samples such as plasma or serum, not the least for clinical purposes. Proteomics keeps generating hundreds of biomarker candidates that need to be transferred towards true clinical application through targeted verification studies and towards clinically applicable analysis formats. The immunoaffinity assay AFFIRM (AFFInity sRM) combines the sensitivity of recombinant single chain antibodies (scFv) for targeted protein enrichment with a specific mass spectrometry readout through selected reaction monitoring (SRM) in an automated workflow. Here we demonstrate a 100 times improved detection capacity of the assay down to pg/ml range... (More)

There is a great need for targeted protein assays with the capacity of sensitive measurements in complex samples such as plasma or serum, not the least for clinical purposes. Proteomics keeps generating hundreds of biomarker candidates that need to be transferred towards true clinical application through targeted verification studies and towards clinically applicable analysis formats. The immunoaffinity assay AFFIRM (AFFInity sRM) combines the sensitivity of recombinant single chain antibodies (scFv) for targeted protein enrichment with a specific mass spectrometry readout through selected reaction monitoring (SRM) in an automated workflow. Here we demonstrate a 100 times improved detection capacity of the assay down to pg/ml range through the use of oriented antibody immobilization to magnetic beads. This was achieved using biotin-tagged scFv coupled to streptavidin coated magnetic beads, or utilizing the FLAG tag for coupling to anti-FLAG antibody coated magnetic beads. An improved multiplexing capacity with an 11-plex setup was also demonstrated compared to a previous 3-plex setup, which is of great importance for the analysis of panels of biomarker targets.

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author
organization
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type
Contribution to journal
publication status
published
subject
in
PLoS ONE
volume
13
issue
2
publisher
Public Library of Science
external identifiers
  • scopus:85042097260
ISSN
1932-6203
DOI
10.1371/journal.pone.0189116
language
English
LU publication?
yes
id
e2e34d95-f44a-47ad-9ce8-69f4430cd75e
date added to LUP
2018-03-07 08:06:18
date last changed
2018-05-29 09:51:56
@article{e2e34d95-f44a-47ad-9ce8-69f4430cd75e,
  abstract     = {<p>There is a great need for targeted protein assays with the capacity of sensitive measurements in complex samples such as plasma or serum, not the least for clinical purposes. Proteomics keeps generating hundreds of biomarker candidates that need to be transferred towards true clinical application through targeted verification studies and towards clinically applicable analysis formats. The immunoaffinity assay AFFIRM (AFFInity sRM) combines the sensitivity of recombinant single chain antibodies (scFv) for targeted protein enrichment with a specific mass spectrometry readout through selected reaction monitoring (SRM) in an automated workflow. Here we demonstrate a 100 times improved detection capacity of the assay down to pg/ml range through the use of oriented antibody immobilization to magnetic beads. This was achieved using biotin-tagged scFv coupled to streptavidin coated magnetic beads, or utilizing the FLAG tag for coupling to anti-FLAG antibody coated magnetic beads. An improved multiplexing capacity with an 11-plex setup was also demonstrated compared to a previous 3-plex setup, which is of great importance for the analysis of panels of biomarker targets.</p>},
  articleno    = {e0189116},
  author       = {Säll, Anna and Corbee, Daniel and Vikström, Sara and Ottosson, Filip and Persson, Helena and Waldemarson, Sofia},
  issn         = {1932-6203},
  language     = {eng},
  month        = {02},
  number       = {2},
  publisher    = {Public Library of Science},
  series       = {PLoS ONE},
  title        = {Advancing the immunoaffinity platform AFFIRM to targeted measurements of proteins in serum in the pg/ml range},
  url          = {http://dx.doi.org/10.1371/journal.pone.0189116},
  volume       = {13},
  year         = {2018},
}