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Microsampling in Targeted Mass Spectrometry-Based Protein Analysis of Low-Abundance Proteins
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Microsampling in Targeted Mass Spectrometry-Based Protein Analysis of Low-Abundance Proteins

Microsampling in Targeted Mass Spectrometry-Based Protein Analysis of Low-Abundance Proteins

DOI:
10.3791/64473-v

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10:21 min

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January 13, 2023

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, , ,
1Department of Pharmacy,University of Oslo

Chapters

  • 00:05Introduction
  • 00:46Preparation of Antibody-Coated Magnetic Beads
  • 03:41Volumetric Absorptive Microsampling and Elution
  • 04:38Digestion of Eluted VAMS
  • 05:30The Capture of Proteotypic ProGRP Using Antibody-Coated Magnetic Beads
  • 07:37Analysis by LC-MS/MS
  • 08:26Results: Analysis of the MS Chromatograms and Base Peak Chromatograms of the Proteotypic Peptide
  • 09:42Conclusion

Summary

자동 번역

자동 번역

A protocol is presented for the determination of low-abundance biomarkers from dried serum samples exemplified with the biomarker progastrin-releasing peptide (ProGRP). Antibody-coated magnetic beads are used for the selective cleanup and enrichment of a proteotypic ProGRP peptide. The captured peptide is subsequently analyzed by liquid chromatography-tandem mass spectrometry.

Tags

MicrosamplingTargeted Mass SpectrometryProtein AnalysisLow-abundance ProteinsAffinity CleanupProtein ProteolysisAntibodyPH AdjustmentMagnetic BeadsAntibody CouplingBorate BufferCoupling Buffer

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