Compatibility of Distinct Label-Free Proteomic Workflows in Absolute Quantification of Proteins Linked to the Oocyte Quality in Human Follicular Fluid
| Autor: | Anna Fel, Marcel Thiel, Paulina Czaplewska, Stanisław Ołdziej, Jacek R. Wiśniewski, Krzysztof Łukaszuk, Aleksandra E. Lewandowska |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Proteomics
0301 basic medicine Swath ms QH301-705.5 Absolute quantification proteome education Pilot Projects Fertilization in Vitro Computational biology Article Catalysis Workflow Inorganic Chemistry 03 medical and health sciences 0302 clinical medicine medicine Humans oocyte quality control Biology (General) Physical and Theoretical Chemistry LC-MS/MS oocyte maturity QD1-999 Molecular Biology Spectroscopy Label free Total protein 030219 obstetrics & reproductive medicine Total Protein Approach Chemistry Organic Chemistry General Medicine blastocyst development Oocyte Follicular fluid Follicular Fluid Computer Science Applications 030104 developmental biology medicine.anatomical_structure Proteome Oocytes SWATH-MS Female Biomarkers human follicular fluid |
| Zdroj: | International Journal of Molecular Sciences Volume 22 Issue 14 International Journal of Molecular Sciences, Vol 22, Iss 7415, p 7415 (2021) |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms22147415 |
| Popis: | We present two separate label-free quantitative workflows based on different high-resolution mass spectrometers and LC setups, which are termed after the utilized instrument: Quad-Orbitrap (nano-LC) and Triple Quad-TOF (micro-LC) and their directed adaptation toward the analysis of human follicular fluid proteome. We identified about 1000 proteins in each distinct workflow using various sample preparation methods. With assistance of the Total Protein Approach, we were able to obtain absolute protein concentrations for each workflow. In a pilot study of twenty samples linked to diverse oocyte quality status from four donors, 455 and 215 proteins were quantified by the Quad-Orbitrap and Triple Quad-TOF workflows, respectively. The concentration values obtained from both workflows correlated to a significant degree. We found reasonable agreement of both workflows in protein fold changes between tested groups, resulting in unified lists of 20 and 22 proteins linked to oocyte maturity and blastocyst development, respectively. The Quad-Orbitrap workflow was best suited for an in-depth analysis without the need of extensive fractionation, especially of low abundant proteome, whereas the Triple Quad-TOF workflow allowed a more robust approach with a greater potential to increase in effectiveness with the growing number of analyzed samples after the initial effort of building a comprehensive spectral library. |
| Databáze: | OpenAIRE |
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