A single microfluidic device for multi-omics analysis sample preparation

• Published in: Lab on a chip Vol. 25; no. 4; pp. 59 - 599
• Main Authors: Kumar, Ranjith Kumar Ravi, Haddad, Iman, Ndiaye, Massamba Mbacké, Marbouty, Martial, Vinh, Joëlle, Verdier, Yann
• Format: Journal Article
• Language: English
• Published: England Royal Society of Chemistry 11.02.2025
• Subjects: Bacillus subtilis - chemistry; Bacillus subtilis - genetics; Chromatography, Liquid; E coli; Escherichia coli - chemistry; Escherichia coli - genetics; Genomics - instrumentation; Lab-On-A-Chip Devices; Microfluidic Analytical Techniques - instrumentation; Microfluidic devices; Microorganisms; Multiomics; Nucleic acids; Proteins; Proteomics; Proteomics - instrumentation; Proteomics - methods; Saccharomyces cerevisiae - chemistry; Saccharomyces cerevisiae - genetics; Samples; Tandem Mass Spectrometry; Workflow
• ISSN: 1473-0197; 1473-0189; 1473-0189
• DOI: 10.1039/d4lc00919c

Combining different "omics" approaches, such as genomics and proteomics, is necessary to generate a detailed and complete insight into microbiome comprehension. Proper sample collection and processing and accurate analytical methods are crucial in generating reliable data. We previously developed the ChipFilter device for proteomic analysis of microbial samples. We have shown that this device coupled to LC-MS/MS can successfully be used to identify microbial proteins. In the present work, we have developed our workflow to analyze concomitantly proteins and nucleic acids from the same sample. We performed lysis and proteolysis in the device using cultures of E. coli , B. subtilis , and S. cerevisiae . After peptide recovery for LC-MS/MS analysis, DNA from the same samples was recovered and successfully amplified by PCR for the 3 species. This workflow was further extended to a complex microbial mixture of known compositions. Protein analysis was carried out, enabling the identification of more than 5000 proteins. The recovered DNA was sequenced, performing comparable to DNA extracted with a commercial kit without proteolysis. Our results show that the ChipFilter device is suited to prepare samples for parallel proteomic and genomic analyses, which is particularly relevant in the case of low-abundant samples and drastically reduces sampling bias. The ChipFilter device is suited to prepare microorganism samples for parallel proteomic and genomic analyses, which is particularly relevant in the case of low-abundant samples and drastically reduces sampling bias.