Your browser doesn't support javascript.
loading
Evaluating the Characteristics of Reporter Ion Signal Acquired in the Orbitrap Analyzer for Isobaric Mass Tag Proteome Quantification Experiments.
Hughes, Christopher S; Zhu, Chenchen; Spicer, Victor; Krokhin, Oleg V; Morin, Gregg B.
Afiliação
  • Hughes CS; Canada's Michael Smith Genome Sciences Centre, British Columbia Cancer Agency , Vancouver, British Columbia V5Z 1L3, Canada.
  • Zhu C; European Molecular Biology Laboratory , Meyerhofstrasse 1, 69117 Heidelberg, Germany.
  • Spicer V; Manitoba Centre for Proteomics and Systems Biology, University of Manitoba , Winnipeg, Manitoba R3E 3P4, Canada.
  • Krokhin OV; Manitoba Centre for Proteomics and Systems Biology, University of Manitoba , Winnipeg, Manitoba R3E 3P4, Canada.
  • Morin GB; Department of Internal Medicine, University of Manitoba , Winnipeg, Manitoba R3A 1R9, Canada.
J Proteome Res ; 16(5): 1831-1838, 2017 05 05.
Article em En | MEDLINE | ID: mdl-28418254
Multiplexed quantification with isobaric chemical tags (e.g., TMT, iTRAQ) provides a robust and efficient means to comparatively examine proteome dynamics between several biological states using a mass spectrometer (MS). The quantitative nature of isobaric tags necessitates strict validation of the observed ion signals in the chosen MS detector before differential patterns are extracted between biological states. We present an in-depth analysis of isobaric tag data acquired on current generation Orbitrap MS hardware to illustrate pitfalls in acquisition settings that can negatively impact results. We establish, for the first time, the presence of a notch, a region of no observed values, in the reporter ion distributions from isobaric-labeled peptide mixtures acquired on these instruments. We determine that this notch is present in published data across a wide range of instruments of the same or different type and is isolated to the Orbitrap mass analyzer. We demonstrate that the impact of the notch can be minimized using manipulations of Orbitrap scan parameters and on-column injection amounts. Lastly, using a mixture of synthetic standard peptides we investigated the impact on identification rates and quantification precision. Together, these data highlight an important phenomenon that negatively impacts peptide identification and quantification in the Orbitrap analyzer as well as outlining guidelines to follow to ensure minimization of MS-induced artifacts in isobaric tag experiments resulting from the notch.
Assuntos
Palavras-chave

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Espectrometria de Massas / Proteoma / Proteômica Tipo de estudo: Prognostic_studies Idioma: En Revista: J Proteome Res Assunto da revista: BIOQUIMICA Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Canadá

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Espectrometria de Massas / Proteoma / Proteômica Tipo de estudo: Prognostic_studies Idioma: En Revista: J Proteome Res Assunto da revista: BIOQUIMICA Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Canadá