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AFFIRM – a multiplexed immunoaffinity platform that combines recombinant antibody fragments and LC-SRM analysis

Säll, Anna LU ; Carlsson, Fredrika LU ; Olsson, Niclas LU ; Wingren, Christer LU ; Ohlin, Mats LU ; Persson, Helena LU and Waldemarson, Sofia LU (2014) In Journal of Proteome Research 13(12). p.5837-5847
Abstract
Targeted measurements of low abundance proteins in complex mixtures are in high demanded in many areas, not the least in clinical applications measuring biomarkers. We here present the novel platform AFFIRM (AFFInity sRM) that utilizes the power of antibody fragments (scFv) to efficiently enrich for target proteins from a complex background and the exquisite specificity of an SRM-MS based detection. To demonstrate the ability of AFFIRM, three target proteins of interest were measured in a serum background in single- and multiplexed experiments in a concentration range of 5-1000 ng/ml. Linear responses were demonstrated down to low ng/ml concentrations with high reproducibility. The platform allows for high throughput measurements in... (More)
Targeted measurements of low abundance proteins in complex mixtures are in high demanded in many areas, not the least in clinical applications measuring biomarkers. We here present the novel platform AFFIRM (AFFInity sRM) that utilizes the power of antibody fragments (scFv) to efficiently enrich for target proteins from a complex background and the exquisite specificity of an SRM-MS based detection. To demonstrate the ability of AFFIRM, three target proteins of interest were measured in a serum background in single- and multiplexed experiments in a concentration range of 5-1000 ng/ml. Linear responses were demonstrated down to low ng/ml concentrations with high reproducibility. The platform allows for high throughput measurements in 96-well format and all steps are amendable to automation and scale-up. We believe the use of recombinant antibody technology in combination with SRM MS analysis provides a powerful way to reach sensitivity, specificity and reproducibility as well as the opportunity to build resources for fast on demand implementation of novel assays. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Proteome Research
volume
13
issue
12
pages
5837 - 5847
publisher
The American Chemical Society
external identifiers
  • wos:000346039400048
  • scopus:84915822551
ISSN
1535-3893
DOI
10.1021/pr500757s
language
English
LU publication?
yes
id
55633650-0e3c-4c85-a407-f4e314fcdea5 (old id 4762096)
date added to LUP
2014-11-06 14:50:20
date last changed
2017-09-03 03:04:16
@article{55633650-0e3c-4c85-a407-f4e314fcdea5,
  abstract     = {Targeted measurements of low abundance proteins in complex mixtures are in high demanded in many areas, not the least in clinical applications measuring biomarkers. We here present the novel platform AFFIRM (AFFInity sRM) that utilizes the power of antibody fragments (scFv) to efficiently enrich for target proteins from a complex background and the exquisite specificity of an SRM-MS based detection. To demonstrate the ability of AFFIRM, three target proteins of interest were measured in a serum background in single- and multiplexed experiments in a concentration range of 5-1000 ng/ml. Linear responses were demonstrated down to low ng/ml concentrations with high reproducibility. The platform allows for high throughput measurements in 96-well format and all steps are amendable to automation and scale-up. We believe the use of recombinant antibody technology in combination with SRM MS analysis provides a powerful way to reach sensitivity, specificity and reproducibility as well as the opportunity to build resources for fast on demand implementation of novel assays.},
  author       = {Säll, Anna and Carlsson, Fredrika and Olsson, Niclas and Wingren, Christer and Ohlin, Mats and Persson, Helena and Waldemarson, Sofia},
  issn         = {1535-3893},
  language     = {eng},
  number       = {12},
  pages        = {5837--5847},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {AFFIRM – a multiplexed immunoaffinity platform that combines recombinant antibody fragments and LC-SRM analysis},
  url          = {http://dx.doi.org/10.1021/pr500757s},
  volume       = {13},
  year         = {2014},
}