Strategy for surveying the proteome using affinity proteomics and mass spectrometry.
Wingren, Christer LU ; James, Peter LU
and Borrebaeck, Carl
LU
(2009)
In Proteomics
9(6).
p.1511-1517
- Abstract
- Antibody-based microarrays is a rapidly evolving technology that has gone from the first proof-of-concept studies to more demanding proteome profiling applications, during the last years. Miniaturized microarrays can be printed with large number of antibodies harbouring predetermined specificities, capable of targeting high- as well as low-abundant analytes in complex, nonfractionated proteomes. Consequently, the resolution of such proteome profiling efforts correlate directly to the number of antibodies included, which today is a key limiting factor. To overcome this bottleneck and to be able to perform in-depth global proteome surveys, we propose to interface affinity proteomics with MS-based read-out, as outlined in this technical... (More)
- Antibody-based microarrays is a rapidly evolving technology that has gone from the first proof-of-concept studies to more demanding proteome profiling applications, during the last years. Miniaturized microarrays can be printed with large number of antibodies harbouring predetermined specificities, capable of targeting high- as well as low-abundant analytes in complex, nonfractionated proteomes. Consequently, the resolution of such proteome profiling efforts correlate directly to the number of antibodies included, which today is a key limiting factor. To overcome this bottleneck and to be able to perform in-depth global proteome surveys, we propose to interface affinity proteomics with MS-based read-out, as outlined in this technical perspective. Briefly, we have defined a range of peptide motifs, each motif being present in 5-100 different proteins. In this manner, 100 antibodies, binding 100 different motifs commonly distributed among different proteins, would potentially target a protein cluster of 10(4) individual molecules, i.e. around 50% of the nonredundant human proteome. Notably, these motif-specific antibodies would be directly applicable to any proteome in a specie independent manner and not biased towards abundant proteins or certain protein classes. The biological sample is digested, exposed to these immobilized antibodies, whereby motif-containing peptides are specifically captured, enriched and subsequently detected and identified using MS. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1302297
- author
- Wingren, Christer
LU
; James, Peter
LU
and Borrebaeck, Carl
LU
- organization
- publishing date
- 2009
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Proteomics
- volume
- 9
- issue
- 6
- pages
- 1511 - 1517
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- wos:000264890400009
- pmid:19235165
- scopus:63049087845
- ISSN
- 1615-9861
- DOI
- 10.1002/pmic.200800802
- language
- English
- LU publication?
- yes
- id
- 545b5200-87c6-4e17-ad3b-6058caff0f91 (old id 1302297)
- date added to LUP
- 2016-04-01 12:25:37
- date last changed
- 2025-10-14 12:28:55
@article{545b5200-87c6-4e17-ad3b-6058caff0f91,
abstract = {{Antibody-based microarrays is a rapidly evolving technology that has gone from the first proof-of-concept studies to more demanding proteome profiling applications, during the last years. Miniaturized microarrays can be printed with large number of antibodies harbouring predetermined specificities, capable of targeting high- as well as low-abundant analytes in complex, nonfractionated proteomes. Consequently, the resolution of such proteome profiling efforts correlate directly to the number of antibodies included, which today is a key limiting factor. To overcome this bottleneck and to be able to perform in-depth global proteome surveys, we propose to interface affinity proteomics with MS-based read-out, as outlined in this technical perspective. Briefly, we have defined a range of peptide motifs, each motif being present in 5-100 different proteins. In this manner, 100 antibodies, binding 100 different motifs commonly distributed among different proteins, would potentially target a protein cluster of 10(4) individual molecules, i.e. around 50% of the nonredundant human proteome. Notably, these motif-specific antibodies would be directly applicable to any proteome in a specie independent manner and not biased towards abundant proteins or certain protein classes. The biological sample is digested, exposed to these immobilized antibodies, whereby motif-containing peptides are specifically captured, enriched and subsequently detected and identified using MS.}},
author = {{Wingren, Christer and James, Peter and Borrebaeck, Carl}},
issn = {{1615-9861}},
language = {{eng}},
number = {{6}},
pages = {{1511--1517}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Proteomics}},
title = {{Strategy for surveying the proteome using affinity proteomics and mass spectrometry.}},
url = {{http://dx.doi.org/10.1002/pmic.200800802}},
doi = {{10.1002/pmic.200800802}},
volume = {{9}},
year = {{2009}},
}