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

Constance A Sobsey; Bjoern Froehlich; Gerald Batist; Christoph H Borchers

doi:10.1007/978-1-0716-2409-8_13

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

Methods Mol Biol. 2022:2515:203-225. doi: 10.1007/978-1-0716-2409-8_13.

Authors

Constance A Sobsey^{1

2}, Bjoern Froehlich^{3

4}, Gerald Batist^{2

5

6}, Christoph H Borchers^{7

8}

Affiliations

¹ Segal Cancer Proteomics Centre, Lady Davis Institute, Jewish General Hospital, McGill University, Montreal, QC, Canada.
² Division of Experimental Medicine, McGill University, Montreal, QC, Canada.
³ University of Victoria - Genome BC Proteomics Centre, Victoria, BC, Canada.
⁴ Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada.
⁵ Gerald Bronfman Department of Oncology, Jewish General Hospital, McGill University, Montreal, QC, Canada.
⁶ Exactis Innovation, Montreal, QC, Canada.
⁷ Segal Cancer Proteomics Centre, Lady Davis Institute, Jewish General Hospital, McGill University, Montreal, QC, Canada. [email protected].
⁸ Gerald Bronfman Department of Oncology, Jewish General Hospital, McGill University, Montreal, QC, Canada. [email protected].

PMID: 35776354
DOI: 10.1007/978-1-0716-2409-8_13

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.

Keywords: Absolute protein quantitation; Immunoenrichment; Internal peptide standard; MALDI; Proteomics; TOF mass spectrometry (MS); iMALDI.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Peptides* / analysis
Proteins* / analysis
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods

Substances

Peptides
Proteins