Use of recombinant proteins as a simple and robust normalization method for untargeted proteomics screening: exhaustive performance assessment

Sandra Isabel Anjo*, Isaura Simões, P. Castanheira, Mário Grãos, B. Manadas

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

The label-free quantitative mass spectrometry methods, in particular, the SWATH-MS approach, have gained popularity and became a powerful technique for comparison of large datasets. In the present work, it is evaluated the use of recombinant proteins as internal standards for untargeted label-free methods. The proposed internal standard strategy reveals a similar intragroup normalization capacity when compared with the most common normalization methods, with the additional advantage of maintaining the overall proteome changes between groups (which is buffered with the use of other methods). Therefore, the proposed strategy is able to maintain a good performance even when large qualitative and quantitative differences in sample composition are observed, such as the ones induced by biological regulation (as observed in secretome and other biofluids’ analyses) or by enrichment approaches (such as immunopurifications). Moreover, this approach corresponds to a cost-effective and simple normalization method altrenative, therefore being an appealing strategy for large quantitative screening, as the analysis of clinical cohorts for biomarker discovery.
Original languageEnglish
Article number120163
JournalTalanta
Volume205
DOIs
Publication statusPublished - 1 Dec 2019
Externally publishedYes

Keywords

  • Biofluids
  • Data normalization
  • Internal standard
  • Proteomics screening
  • SWATH-MS

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