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Mass-Tag Enhanced Immuno-Laser Desorption/Ionization Mass Spectrometry for Sensitive Detection of Intact Protein Antigens

Lorey, Martina; Adler, Belinda LU ; Yan, Hong LU ; Soliymani, Rabah; Ekström, Simon LU ; Yli-Kauhaluoma, Jari; Laurell, Thomas LU and Baumann, Marc (2015) In Analytical Chemistry 87(10). p.5255-5262
Abstract
A new read-out method for antibody arrays using laser desorption/ionization-mass spectrometry (LDI-MS) is presented. Small, photocleavable reporter molecules with a defined mass called "mass-tags" are used for detection of immunocaptured proteins from human plasma. Using prostate specific antigen (PSA), a biomarker for prostate cancer, as a model antigen, a high sensitivity generic detection methodology based immunocapture with a primary antibody and with a biotin labeled secondary antibody coupled to mass-tagged avidin is demonstrated. As each secondary antibody can bind several avidin molecules, each having a large number of mass-tags, signal amplification can be achieved. The developed PSA sandwich mass-tag analysis method provided a... (More)
A new read-out method for antibody arrays using laser desorption/ionization-mass spectrometry (LDI-MS) is presented. Small, photocleavable reporter molecules with a defined mass called "mass-tags" are used for detection of immunocaptured proteins from human plasma. Using prostate specific antigen (PSA), a biomarker for prostate cancer, as a model antigen, a high sensitivity generic detection methodology based immunocapture with a primary antibody and with a biotin labeled secondary antibody coupled to mass-tagged avidin is demonstrated. As each secondary antibody can bind several avidin molecules, each having a large number of mass-tags, signal amplification can be achieved. The developed PSA sandwich mass-tag analysis method provided a limit of detection below 200 pg/mL (6 pM) for a 10 mu L plasma sample, Well below the clinically relevant cutoff value of 3-4 ng/mL. This brings the limit of detection (LOD) for detection of intact antigens with matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) down to levels comparable to capture by anti-peptide antibodies selected reaction monitoring (SISCAPA SRM) and enzyme linked immunosorbent assay (ELISA), as 6 pM corresponds to a maximal amount of 60 amol PSA captured on-spot. We :propose the potential use Of LDI (laser desorption/ionization) with mass-tag read-out implemented in a sandwich assay format for low abundant and/or early disease biomarker detection. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Chemistry
volume
87
issue
10
pages
5255 - 5262
publisher
The American Chemical Society
external identifiers
  • wos:000355057700034
  • scopus:84929598060
ISSN
1520-6882
DOI
10.1021/acs.analchem.5b00304
language
English
LU publication?
yes
id
0b3ad21f-2cf1-4cd4-889d-b5bedcb3e472 (old id 7410684)
date added to LUP
2015-06-29 11:16:54
date last changed
2017-04-09 03:17:54
@article{0b3ad21f-2cf1-4cd4-889d-b5bedcb3e472,
  abstract     = {A new read-out method for antibody arrays using laser desorption/ionization-mass spectrometry (LDI-MS) is presented. Small, photocleavable reporter molecules with a defined mass called "mass-tags" are used for detection of immunocaptured proteins from human plasma. Using prostate specific antigen (PSA), a biomarker for prostate cancer, as a model antigen, a high sensitivity generic detection methodology based immunocapture with a primary antibody and with a biotin labeled secondary antibody coupled to mass-tagged avidin is demonstrated. As each secondary antibody can bind several avidin molecules, each having a large number of mass-tags, signal amplification can be achieved. The developed PSA sandwich mass-tag analysis method provided a limit of detection below 200 pg/mL (6 pM) for a 10 mu L plasma sample, Well below the clinically relevant cutoff value of 3-4 ng/mL. This brings the limit of detection (LOD) for detection of intact antigens with matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) down to levels comparable to capture by anti-peptide antibodies selected reaction monitoring (SISCAPA SRM) and enzyme linked immunosorbent assay (ELISA), as 6 pM corresponds to a maximal amount of 60 amol PSA captured on-spot. We :propose the potential use Of LDI (laser desorption/ionization) with mass-tag read-out implemented in a sandwich assay format for low abundant and/or early disease biomarker detection.},
  author       = {Lorey, Martina and Adler, Belinda and Yan, Hong and Soliymani, Rabah and Ekström, Simon and Yli-Kauhaluoma, Jari and Laurell, Thomas and Baumann, Marc},
  issn         = {1520-6882},
  language     = {eng},
  number       = {10},
  pages        = {5255--5262},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Mass-Tag Enhanced Immuno-Laser Desorption/Ionization Mass Spectrometry for Sensitive Detection of Intact Protein Antigens},
  url          = {http://dx.doi.org/10.1021/acs.analchem.5b00304},
  volume       = {87},
  year         = {2015},
}