Immuno-MALDI-MS for Accurate Quantitation of Targeted Peptides from Volume-Restricted Samples

Constance A. Sobsey, Bjoern Froehlich, Gerald Batist, Christoph H. Borchers

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

The immuno-MALDI-MS method can be used to quantify low-abundance proteins from clinical samples that offer only a limited amount of material for analysis. An internal standard, in the form of a stable isotope-labeled peptide, is used to ensure reproducible and absolute quantitation. The protocol described here was optimized for the quantitation of AKT1 and AKT2, but we offer instructions on how to adapt the method to target other proteins. The described workflow is compatible with automation via a liquid handling robot for high-throughput applications.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages203-225
Number of pages23
DOIs
Publication statusPublished - 2022
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2515
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Absolute protein quantitation
  • iMALDI
  • Immunoenrichment
  • Internal peptide standard
  • MALDI
  • Proteomics
  • TOF mass spectrometry (MS)

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