Comprehensive Micro-SPE-Based Bottom-Up Proteomic Workflow for Sensitive Analysis of Limited Samples.
Methods Mol Biol
; 2817: 19-31, 2024.
Article
em En
| MEDLINE
| ID: mdl-38907144
ABSTRACT
Clinical and biological samples are often scarce and precious (e.g., rare cell isolates, microneedle tissue biopsies, small-volume liquid biopsies, and even single cells or organelles). Typical large-scale proteomic methods, where significantly higher protein amounts are analyzed, are not directly transferable to the analysis of limited samples due to their incompatibility with pg-, ng-, and low-µg-level protein sample amounts. Here, we report the on-microsolid-phase extraction tip (OmSET)-based sample preparation workflow for sensitive analysis of limited biological samples to address this challenge. The developed platform was successfully tested for the analysis of 100-10,000 typical mammalian cells and is scalable to allow for lower and larger protein amounts and more samples to be analyzed (i.e., higher throughput of analysis).
Palavras-chave
BUP (bottom-up proteomics); DIA (data-independent acquisition); Limited samples; MS (mass spectrometry); Micro-SPE (micro-solid phase extraction); OmSET (on-microsolid-phase extraction tip); On-membrane digestion; Sample preparation; nanoLC-MS (nanoflow liquid chromatography coupled to mass spectrometry)
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteômica
/
Extração em Fase Sólida
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Fluxo de Trabalho
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article