Journal
/
Chemistry
/
Microsampling in Targeted Mass Spectrometry-Based Protein Analysis of Low-Abundance Proteins
JoVE Journal
Chemistry
This content is Free Access.
JoVE Journal Chemistry
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

•

10:21 min

•

January 13, 2023

•

, , ,
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

Automatic Translation

Automatic Translation

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

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/64473?article_type=v

Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)
check_url/64473?article_type=v

Matrix-assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) Mass Spectrometric Analysis of Intact Proteins Larger than 100 kDa
check_url/64473?article_type=v

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
check_url/64473?article_type=v

Mass Spectrometric Approaches to Study Protein Structure and Interactions in Lyophilized Powders
check_url/64473?article_type=v

Sequencing of Plant Wall Heteroxylans Using Enzymic, Chemical (Methylation) and Physical (Mass Spectrometry, Nuclear Magnetic Resonance) Techniques
check_url/64473?article_type=v

An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry
check_url/64473?article_type=v

Real-time Breath Analysis by Using Secondary Nanoelectrospray Ionization Coupled to High Resolution Mass Spectrometry
check_url/64473?article_type=v

Synthesis and Mass Spectrometry Analysis of Oligo-peptoids
check_url/64473?article_type=v

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry
check_url/64473?article_type=v

Directed Assembly of Elastin-like Proteins into defined Supramolecular Structures and Cargo Encapsulation In Vitro

Read Article