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An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry
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Chemie
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JoVE Journal Chemie
An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry

An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry

DOI:
10.3791/55325-v

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11:09 min

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April 17, 2018

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, , , , ,
1Waters Corporation

Kapitel

  • 00:04Titel
  • 00:44Setup of the LC/HDMSE (High Definition MSE) Data Acquisition Method
  • 03:59Setup of the Tof-MRM Method for Collision Energy Optimization and the Final HS-MRM Acquisition Method for Peptide Quantification
  • 07:42Creation of a Processing Method for the Analysis of the HS-MRM Dataset
  • 09:57Results: An HS-MRM Assay for Quantification of Host-cell Protein Peptides in a Monoclonal Antibody Digest
  • 10:29Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

Here, we describe a chromatographic assay coupled with the ion mobility separation of peptide precursors followed by the high-resolution (~30,000) MS-detection of peptide fragments for the quantification of spiked peptide standards in a monoclonal antibody digest.

Tags

Keywords: HS-MRM AssayHost-cell ProteinsProtein BiopharmaceuticalsLiquid ChromatographyIon MobilityQTOF Mass SpectrometryPeptide QuantificationProtein CharacterizationQuantitative ProteomicsLow-abundant ProteinsPeptide SeparationGradient ElutionIon Mobility SeparationHigh-resolution Mass SpectrometryHigh-definition MSECollision EnergyLock Mass Correction

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