Benchmarking accuracy and precision of intensity-based absolute quantification of protein abundances in Saccharomyces cerevisiae
Artikel i vetenskaplig tidskrift, 2021
Protein quantification via label-free mass spectrometry (MS) has become an increasingly popular method for predicting genome-wide absolute protein abundances. A known caveat of this approach, however, is the poor technical reproducibility, that is, how consistent predictions are when the same sample is measured repeatedly. Here, we measured proteomics data for Saccharomyces cerevisiae with both biological and inter-batch technical triplicates, to analyze both accuracy and precision of protein quantification via MS. Moreover, we analyzed how these metrics vary when applying different methods for converting MS intensities to absolute protein abundances. We demonstrate that our simple normalization and rescaling approach can perform as accurately, yet more precisely, than methods which rely on external standards. Additionally, we show that inter-batch reproducibility is worse than biological reproducibility for all evaluated methods. These results offer a new benchmark for assessing MS data quality for protein quantification, while also underscoring current limitations in this approach.
UPS2
mass spectrometry
batch effect
absolute proteomics
iBAQ
Författare
Benjamín José Sánchez
Chalmers, Biologi och bioteknik, Systembiologi
Novo Nordisk Foundation Center for Biosustainability
Petri-Jaan Lahtvee
Tartu Ülikool
Kate Campbell
Chalmers, Biologi och bioteknik, Systembiologi
Novo Nordisk Foundation Center for Biosustainability
S. Kasvandik
Tartu Ülikool
Tao Yu
Novo Nordisk Foundation Center for Biosustainability
Chalmers, Biologi och bioteknik, Systembiologi
Iván Domenzain Del Castillo Cerecer
Novo Nordisk Foundation Center for Biosustainability
Chalmers, Biologi och bioteknik, Systembiologi
Aleksej Zelezniak
Chalmers, Biologi och bioteknik, Systembiologi
Jens B Nielsen
Novo Nordisk Foundation Center for Biosustainability
Chalmers, Biologi och bioteknik, Systembiologi
Danmarks Tekniske Universitet (DTU)
Proteomics
1615-9853 (ISSN) 1615-9861 (eISSN)
Vol. In PressÄmneskategorier
Analytisk kemi
Bioinformatik (beräkningsbiologi)
Bioinformatik och systembiologi
DOI
10.1002/pmic.202000093
PubMed
33452728