Digital Microfluidics for Microproteomic Analysis of Minute Mammalian Tissue Samples Enabled by a Photocleavable Surfactant.

Autor:	Chan C; Department of Chemistry, University of Toronto, Toronto M5S 3H6, Ontario, Canada., Peng J; Department of Chemistry, University of Toronto, Toronto M5S 3H6, Ontario, Canada.; Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto M5S 3E1, Ontario, Canada.; Institute of Biomedical Engineering, University of Toronto, Toronto M5S 3G9, Ontario, Canada., Rajesh V; Department of Chemistry, University of Toronto, Toronto M5S 3H6, Ontario, Canada., Scott EY; Department of Chemistry, University of Toronto, Toronto M5S 3H6, Ontario, Canada.; Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto M5S 3E1, Ontario, Canada.; Institute of Biomedical Engineering, University of Toronto, Toronto M5S 3G9, Ontario, Canada.; Department of Surgery, University of Toronto, Toronto M5S 1A8, Ontario, Canada., Sklavounos AA; Department of Chemistry, University of Toronto, Toronto M5S 3H6, Ontario, Canada.; Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto M5S 3E1, Ontario, Canada., Faiz M; Department of Surgery, University of Toronto, Toronto M5S 1A8, Ontario, Canada., Wheeler AR; Department of Chemistry, University of Toronto, Toronto M5S 3H6, Ontario, Canada.; Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto M5S 3E1, Ontario, Canada.; Institute of Biomedical Engineering, University of Toronto, Toronto M5S 3G9, Ontario, Canada.
Jazyk:	angličtina
Zdroj:	Journal of proteome research [J Proteome Res] 2023 Oct 06; Vol. 22 (10), pp. 3242-3253. Date of Electronic Publication: 2023 Aug 31.
DOI:	10.1021/acs.jproteome.3c00281
Abstrakt:	Proteome profiles of precious tissue samples have great clinical potential for accelerating disease biomarker discovery and promoting novel strategies for early diagnosis and treatment. However, tiny clinical tissue samples are often difficult to handle and analyze with conventional proteomic methods. Automated digital microfluidic (DMF) workflows facilitate the manipulation of size-limited tissue samples. Here, we report the assessment of a DMF microproteomics workflow enabled by a photocleavable surfactant for proteomic analysis of minute tissue samples. The surfactant 4-hexylphenylazosulfonate (Azo) was found to facilitate fast droplet movement on DMF and enhance the proteomics analysis. Comparisons of Azo and n -Dodecyl β-d-maltoside (DDM) using small samples of HeLa digest standards and MCF-7 cell digests revealed distinct differences at the peptide level despite similar results at the protein level. The DMF microproteomics workflow was applied for the sample preparation of ∼3 μg biopsies from murine brain tissue. A total of 1969 proteins were identified in three samples, including established neural biomarkers and proteins related to synaptic signaling. Going forward, we propose that the Azo-enabled DMF workflow has the potential to advance the practical clinical application of DMF for the analysis of size-limited tissue samples.
Databáze:	MEDLINE
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