Advancing Nanomaterial Toxicology Screening Through Efficient and Cost‐Effective Quantitative Proteomics

Abstract: Proteomic investigations yield high‐dimensional datasets, yet their application to large‐scale toxicological assessments is hindered by reproducibility challenges due to fluctuating measurement conditions. To address these limitations, this study introduces an advanced tandem mass tag (TMT) labeling protocol. Although labeling approaches shorten data acquisition time by multiplexing samples compared to traditional label‐free quantification (LFQ) methods in general, the associated costs may surge significantly with large sample sets, for example, in toxicological screenings. However, the introduced advanced protocol offers an efficient, cost‐effective alternative, reducing TMT reagent usage (by a factor of ten) and requiring minimal biological material (1 µg), while demonstrating increased reproducibility compared to LFQ. To demonstrate its effectiveness, the advanced protocol is employed to assess the toxicity of nine benchmark nanomaterials (NMs) on A549 lung epithelial cells. While LFQ measurements identify 3300 proteins, they proved inadequate to reveal NM toxicity. Conversely, despite detecting 2600 proteins, the TMT protocol demonstrates superior sensitivity by uncovering alterations induced by NM treatment. In contrast to previous studies, the introduced advanced protocol allows simultaneous and straightforward assessment of multiple test substances, enabling prioritization, ranking, and grouping for hazard evaluation. Additionally, it fosters the development of New Approach Methodologies (NAMs), contributing to innovative methodologies in toxicological research.

Location
Deutsche Nationalbibliothek Frankfurt am Main
Extent
Online-Ressource
Language
Englisch

Bibliographic citation
Advancing Nanomaterial Toxicology Screening Through Efficient and Cost‐Effective Quantitative Proteomics ; day:30 ; month:05 ; year:2024 ; extent:11
Small Methods ; (30.05.2024) (gesamt 11)

Creator
Stobernack, Tobias
Dommershausen, Nils
Alcolea‐Rodríguez, Víctor
Ledwith, Rico
Bañares, Miguel A.
Haase, Andrea
Pink, Mario
Dumit, Verónica I.

DOI
10.1002/smtd.202400420
URN
urn:nbn:de:101:1-2405301426198.272197602189
Rights
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
Last update
14.08.2025, 10:57 AM CEST

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Associated

  • Stobernack, Tobias
  • Dommershausen, Nils
  • Alcolea‐Rodríguez, Víctor
  • Ledwith, Rico
  • Bañares, Miguel A.
  • Haase, Andrea
  • Pink, Mario
  • Dumit, Verónica I.

Other Objects (12)