Strategy for surveying the proteome using affinity proteomics and mass spectrometry. Export

@article{citeulike:4154050, 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, C. and James, P. and Borrebaeck, C. A.}, citeulike-article-id = {4154050}, citeulike-linkout-0 = {http://dx.doi.org/10.1002/pmic.200800802}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/19235165}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=19235165}, day = {20}, doi = {10.1002/pmic.200800802}, issn = {1615-9861}, journal = {Proteomics}, keywords = {affinity, ms, s}, month = {March}, number = {6}, pages = {1511--1517}, posted-at = {2009-03-09 04:34:27}, priority = {2}, title = {Strategy for surveying the proteome using affinity proteomics and mass spectrometry.}, url = {http://dx.doi.org/10.1002/pmic.200800802}, volume = {9}, year = {2009} }

TY - JOUR ID - citeulike:4154050 L3 - citeulike-article-id:4154050 N2 - 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. IS - 6 JF - Proteomics SN - 1615-9861 EP - 1517 TI - Strategy for surveying the proteome using affinity proteomics and mass spectrometry. VL - 9 SP - 1511 KW - affinity KW - ms KW - s AU - Wingren, C AU - James, P AU - Borrebaeck, CA PY - 2009/03/20/ UR - http://dx.doi.org/10.1002/pmic.200800802 ER -