Label-Free Quantitation and Mapping of the ErbB2 Tumor Receptor by Multiple Protease Digestion with Data-Dependent (MS1) and Data-Independent (MS2) Acquisitions
Joint Authors
D'Souza, Alexandria K.
Behring, Jessica B.
Gibson, Bradford W.
Schilling, Birgit
Srinivasan, Tara
Benz, Christopher C.
Sorensen, Dylan J.
Held, Jason M.
Source
International Journal of Proteomics
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-04-04
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Natural & Life Sciences (Multidisciplinary)
Biology
Abstract EN
The receptor tyrosine kinase ErbB2 is a breast cancer biomarker whose posttranslational modifications (PTMs) are a key indicator of its activation.
Quantifying the expression and PTMs of biomarkers such as ErbB2 by selected reaction monitoring (SRM) mass spectrometry has several limitations, including minimal coverage and extensive assay development time.
Therefore, we assessed the utility of two high resolution, full scan mass spectrometry approaches, MS1 Filtering and SWATH MS2, for targeted ErbB2 proteomics.
Endogenous ErbB2 immunoprecipitated from SK-BR-3 cells was in-gel digested with trypsin, chymotrypsin, Asp-N, or trypsin plus Asp-N in triplicate.
Data-dependent acquisition with an AB SCIEX TripleTOF 5600 and MS1 Filtering data processing was used to assess peptide and PTM coverage as well as the reproducibility of enzyme digestion.
Data-independent acquisition (SWATH) was also performed for MS2 quantitation.
MS1 Filtering and SWATH MS2 allow quantitation of all detected analytes after acquisition, enabling the use of multiple proteases for quantitative assessment of target proteins.
Combining high resolution proteomics with multiprotease digestion enabled quantitative mapping of ErbB2 with excellent reproducibility, improved amino acid sequence and PTM coverage, and decreased assay development time compared to typical SRM assays.
These results demonstrate that high resolution quantitative proteomic approaches are an effective tool for targeted biomarker quantitation.
American Psychological Association (APA)
Held, Jason M.& Schilling, Birgit& D'Souza, Alexandria K.& Srinivasan, Tara& Behring, Jessica B.& Sorensen, Dylan J.…[et al.]. 2013. Label-Free Quantitation and Mapping of the ErbB2 Tumor Receptor by Multiple Protease Digestion with Data-Dependent (MS1) and Data-Independent (MS2) Acquisitions. International Journal of Proteomics،Vol. 2013, no. 2013, pp.1-11.
https://search.emarefa.net/detail/BIM-498450
Modern Language Association (MLA)
Held, Jason M.…[et al.]. Label-Free Quantitation and Mapping of the ErbB2 Tumor Receptor by Multiple Protease Digestion with Data-Dependent (MS1) and Data-Independent (MS2) Acquisitions. International Journal of Proteomics No. 2013 (2013), pp.1-11.
https://search.emarefa.net/detail/BIM-498450
American Medical Association (AMA)
Held, Jason M.& Schilling, Birgit& D'Souza, Alexandria K.& Srinivasan, Tara& Behring, Jessica B.& Sorensen, Dylan J.…[et al.]. Label-Free Quantitation and Mapping of the ErbB2 Tumor Receptor by Multiple Protease Digestion with Data-Dependent (MS1) and Data-Independent (MS2) Acquisitions. International Journal of Proteomics. 2013. Vol. 2013, no. 2013, pp.1-11.
https://search.emarefa.net/detail/BIM-498450
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-498450