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Related Concept Videos

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Quantitative Magnetic Resonance Imaging of Skeletal Muscle Disease
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SWATH-MS Protocols in Human Diseases.

Maria Del Pilar Chantada-Vázquez1, María García Vence2, Antonio Serna3

  • 1Research Unit, Hospital Universitario Lucus Augusti (HULA), Servizo Galego de Saúde (SERGAS), Lugo, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|March 9, 2021
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Summary

This study details a micro liquid chromatography-sequential window acquisition of all theoretical mass spectra (LC-SWATH-MS) protocol for identifying and quantifying disease biomarkers in biological fluids. This method enhances disease diagnosis and treatment through advanced proteomics.

Keywords:
Biological fluidsLabel-free quantificationNanoproteomicsSWATH-MSSalivaSerum/plasmaTearsUrine

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Area of Science:

  • Translational Science
  • Biomarker Discovery
  • Proteomics

Background:

  • Identifying molecular biomarkers is crucial for understanding human disease origin, progression, diagnosis, prognosis, and treatment.
  • Proteomics offers solutions for analyzing complex biological samples like organic fluids.
  • Advancements in high-throughput, label-free quantitative proteomics (SWATH-MS) and liquid chromatography enable deep analysis of large sample sets.

Purpose of the Study:

  • To present optimized sample processing methods for biological fluids.
  • To demonstrate a micro liquid chromatography-SWATH-MS protocol for protein identification and quantification.
  • To facilitate biomarker discovery in accessible human samples.

Main Methods:

  • Utilized advanced sample processing techniques, including major protein depletion and enzymatic digestion.
  • Employed micro liquid chromatography coupled with SWATH-MS (LC-SWATH-MS) for high-throughput, label-free quantitative proteomics.
  • Applied the protocol to various biological fluid samples: serum/plasma, saliva, urine, and tears.

Main Results:

  • Successfully identified and quantified multiple proteins across different human biofluid types.
  • Demonstrated the sensitivity and depth required for comprehensive proteomic analysis in complex samples.
  • Validated the efficacy of the developed LC-SWATH-MS protocol for biomarker discovery.

Conclusions:

  • The described sample processing and micro LC-SWATH-MS protocol are effective for identifying and quantifying protein biomarkers in human biofluids.
  • This approach supports advancements in translational science for improved disease diagnosis and treatment strategies.
  • The methodology provides a robust platform for large-scale proteomic studies using easily accessible samples.