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Multiple and Selective Reaction Monitoring Using Triple Quadrupole Mass Spectrometer: Preclinical Large Cohort Analysis

  • Qin Fu
  • Zhaohui Chen
  • Shenyan Zhang
  • Sarah J. Parker
  • Zongming Fu
  • Adrienne Tin
  • Xiaoqian Liu
  • Jennifer E. Van Eyk
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1410)

Abstract

Multiple reaction monitoring (MRM), sometimes referred to as selective reaction monitoring (SRM), is a mass spectrometry method that can target selective peptides for the detection and quantitation of a protein. Compared to traditional ELISA, MRM assays have a number of advantages including ease in multiplexing several proteins in the same assay and independence from the necessity for high-quality, expensive, and at times unreliable antibodies. Furthermore, MRM assays can be developed to quantify multiple proteoforms of a single protein allowing the quantification of allelic expression of a particular sequence polymorphism, protein isoform, as well as determining site occupancy of posttranslational modification(s). In this chapter, we describe our workflow for target peptide selection, assay optimization, and acquisition multiplexing. Our workflow is presented using the example of constrained MRM assays developed for the serum protein ApoL1 in its various proteoforms to highlight the specific technical considerations necessary for the difficult task of quantifying peptide targets based on highly specific amino acid sequences by MRM.

Key words

Quantification Multiple reaction monitoring Selective reaction monitoring Mass spectrometry APO L1 

Notes

Acknowledgements

This work was supported by NHLBI Johns Hopkins Proteomic Innovation Center in Heart Failure—HHSN268201000032C (JVE)—and partially supported by the Chronic Kidney Disease Biomarker Consortium funded by NIDDK U01-U01DK085689. Special thanks to Drs. Josef Coresh, Lesley Inker, Chi-yuan Hsu, John Eckfeldt, Paul Kimmel, Dr. Vasan Ramachandran, and Harold I. Feldman.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Qin Fu
    • 1
  • Zhaohui Chen
    • 1
  • Shenyan Zhang
    • 1
  • Sarah J. Parker
    • 1
  • Zongming Fu
    • 2
  • Adrienne Tin
    • 3
  • Xiaoqian Liu
    • 1
  • Jennifer E. Van Eyk
    • 1
  1. 1.Advanced Clinical Biosystems Research Institute, The Heart InstituteCedars-Sinai Medical CenterLos AngelesUSA
  2. 2.Department of MedicineJohns Hopkins UniversityBaltimoreUSA
  3. 3.Department of EpidemiologyJohns Hopkins UniversityBaltimoreUSA

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