Mass-Tag Enhanced Immuno-Laser Desorption/Ionization Mass Spectrometry for Sensitive Detection of Intact Protein Antigens
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/7410684
- author
- Lorey, Martina ; Adler, Belinda LU ; Yan, Hong LU ; Soliymani, Rabah ; Ekström, Simon LU ; Yli-Kauhaluoma, Jari ; Laurell, Thomas LU and Baumann, Marc
- organization
- publishing date
- 2015
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Analytical Chemistry
- volume
- 87
- issue
- 10
- pages
- 5255 - 5262
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000355057700034
- scopus:84929598060
- pmid:25867450
- 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
- 2016-04-01 10:45:28
- date last changed
- 2022-03-04 22:34:44
@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 (ACS)}}, 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}}, doi = {{10.1021/acs.analchem.5b00304}}, volume = {{87}}, year = {{2015}}, }