Résumé
We performed a test sample study to try to identify errors leading to irreproducibility, including incompleteness of peptide sampling, in liquid chromatography-mass spectrometry-based proteomics. We distributed an equimolar test sample, comprising 20 highly purified recombinant human proteins, to 27 laboratories. Each protein contained one or more unique tryptic peptides of 1,250 Da to test for ion selection and sampling in the mass spectrometer. Of the 27 labs, members of only 7 labs initially reported all 20 proteins correctly, and members of only 1 lab reported all tryptic peptides of 1,250 Da. Centralized analysis of the raw data, however, revealed that all 20 proteins and most of the 1,250 Da peptides had been detected in all 27 labs. Our centralized analysis determined missed identifications (false negatives), environmental contamination, database matching and curation of protein identifications as sources of problems. Improved search engines and databases are needed for mass spectrometry-based proteomics.
langue originale | Anglais |
---|---|
Pages (de - à) | 423-430 |
Nombre de pages | 8 |
journal | Nature Methods |
Volume | 6 |
Numéro de publication | 6 |
Les DOIs | |
état | Publié - 18 mai 2009 |
Modification externe | Oui |
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Dans: Nature Methods, Vol 6, Numéro 6, 18.05.2009, p. 423-430.
Résultats de recherche: Contribution à un journal › Article › Revue par des pairs
TY - JOUR
T1 - A HUPO test sample study reveals common problems in mass spectrometry-based proteomics
AU - Bell, Alexander W.
AU - Deutsch, Eric W.
AU - Au, Catherine E.
AU - Kearney, Robert E.
AU - Beavis, Ron
AU - Sechi, Salvatore
AU - Nilsson, Tommy
AU - Bergeron, John J.M.
AU - Beardslee, Thomas A.
AU - Chappell, Thomas
AU - Meredith, Gavin
AU - Sheffield, Peter
AU - Gray, Phillip
AU - Hajivandi, Mahbod
AU - Pope, Marshall
AU - Predki, Paul
AU - Kullolli, Majlinda
AU - Hincapie, Marina
AU - Hancock, William S.
AU - Jia, Wei
AU - Song, Lina
AU - Li, Lei
AU - Wei, Junying
AU - Yang, Bing
AU - Wang, Jinglan
AU - Ying, Wantao
AU - Zhang, Yangjun
AU - Cai, Yun
AU - Qian, Xiaohong
AU - He, Fuchu
AU - Meyer, Helmut E.
AU - Stephan, Christian
AU - Eisenacher, Martin
AU - Marcus, Katrin
AU - Langenfeld, Elmar
AU - May, Caroline
AU - Carr, Steve A.
AU - Ahmad, Rushdy
AU - Zhu, Wenhong
AU - Smith, Jeffrey W.
AU - Hanash, Samir M.
AU - Struthers, Jason J.
AU - Wang, Hong
AU - Zhang, Qing
AU - An, Yanming
AU - Goldman, Radoslav
AU - Carlsohn, Elisabet
AU - van der Post, Sjoerd
AU - Hung, Kenneth E.
AU - Sarracino, David A.
AU - Parker, Kenneth
AU - Krastins, Bryan
AU - Kucherlapati, Raju
AU - Bourassa, Sylvie
AU - Poirier, Guy G.
AU - Kapp, Eugene
AU - Patsiouras, Heather
AU - Moritz, Robert
AU - Simpson, Richard
AU - Houle, Benoit
AU - LaBoissiere, Sylvie
AU - Metalnikov, Pavel
AU - Nguyen, Vivian
AU - Pawson, Tony
AU - Wong, Catherine C.L.
AU - Cociorva, Daniel
AU - Yates, John R.
AU - Ellison, Michael J.
AU - Lopez-Campistrous, Ana
AU - Semchuk, Paul
AU - Wang, Yueju
AU - Ping, Peipei
AU - Elia, Giuliano
AU - Dunn, Michael J.
AU - Wynne, Kieran
AU - Walker, Angela K.
AU - Strahler, John R.
AU - Andrews, Philip C.
AU - Hood, Brian L.
AU - Bigbee, William L.
AU - Conrads, Thomas P.
AU - Smith, Derek
AU - Borchers, Christoph H.
AU - Lajoie, Gilles A.
AU - Bendall, Sean C.
AU - Speicher, Kaye D.
AU - Speicher, David W.
AU - Fujimoto, Masanori
AU - Nakamura, Kazuyuki
AU - Paik, Young Ki
AU - Cho, Sang Yun
AU - Kwon, Min Seok
AU - Lee, Hyoung Joo
AU - Jeong, Seul Ki
AU - Chung, An Sung
AU - Miller, Christine A.
AU - Grimm, Rudolf
AU - Williams, Katy
AU - Dorschel, Craig
AU - Falkner, Jayson A.
AU - Martens, Lennart
AU - Vizcaíno, Juan Antonio
PY - 2009/5/18
Y1 - 2009/5/18
N2 - We performed a test sample study to try to identify errors leading to irreproducibility, including incompleteness of peptide sampling, in liquid chromatography-mass spectrometry-based proteomics. We distributed an equimolar test sample, comprising 20 highly purified recombinant human proteins, to 27 laboratories. Each protein contained one or more unique tryptic peptides of 1,250 Da to test for ion selection and sampling in the mass spectrometer. Of the 27 labs, members of only 7 labs initially reported all 20 proteins correctly, and members of only 1 lab reported all tryptic peptides of 1,250 Da. Centralized analysis of the raw data, however, revealed that all 20 proteins and most of the 1,250 Da peptides had been detected in all 27 labs. Our centralized analysis determined missed identifications (false negatives), environmental contamination, database matching and curation of protein identifications as sources of problems. Improved search engines and databases are needed for mass spectrometry-based proteomics.
AB - We performed a test sample study to try to identify errors leading to irreproducibility, including incompleteness of peptide sampling, in liquid chromatography-mass spectrometry-based proteomics. We distributed an equimolar test sample, comprising 20 highly purified recombinant human proteins, to 27 laboratories. Each protein contained one or more unique tryptic peptides of 1,250 Da to test for ion selection and sampling in the mass spectrometer. Of the 27 labs, members of only 7 labs initially reported all 20 proteins correctly, and members of only 1 lab reported all tryptic peptides of 1,250 Da. Centralized analysis of the raw data, however, revealed that all 20 proteins and most of the 1,250 Da peptides had been detected in all 27 labs. Our centralized analysis determined missed identifications (false negatives), environmental contamination, database matching and curation of protein identifications as sources of problems. Improved search engines and databases are needed for mass spectrometry-based proteomics.
UR - http://www.scopus.com/inward/record.url?scp=67349196123&partnerID=8YFLogxK
U2 - 10.1038/nmeth.1333
DO - 10.1038/nmeth.1333
M3 - Article
C2 - 19448641
AN - SCOPUS:67349196123
SN - 1548-7091
VL - 6
SP - 423
EP - 430
JO - Nature Methods
JF - Nature Methods
IS - 6
ER -