An automated assay for the clinical measurement of plasma renin activity by immuno-MALDI (iMALDI)

Robert Popp, David Malmström, Andrew G. Chambers, David Lin, Alexander G. Camenzind, J. Grace Van Der Gugten, Daniel T. Holmes, Michael Pugia, Marta Jaremek, Shannon Cornett, Detlev Suckau, Christoph H. Borchers

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

Abstract

Plasma renin activity (PRA) is essential for the screening and diagnosis of primary aldosteronism (PA), a form of secondary hypertension, which affects approximately 100 million people worldwide. It is commonly determined by radioimmunoassay (RIA) and, more recently, by relatively low-throughput LC-MS/MS methods. In order to circumvent the negative aspects of RIAs (radioisotopes, cross-reactivity) and the low throughput of LC-MS based methods, we have developed a high-throughput immuno-MALDI (iMALDI)-based assay for PRA determination using an Agilent Bravo for automated liquid handling and a Bruker Microflex LRF instrument for MALDI analysis, with the goal of implementing the assay in clinical laboratories. The current assay allows PRA determination of 29 patient samples (192 immuno-captures), within ~ 6 to 7 h, using a 3-hour Ang I generation period, at a 7.5-fold faster analysis time than LC-MS/MS. The assay is performed on 350 μL of plasma, and has a linear range from 0.08 to 5.3 ng/L/s in the reflector mode, and 0.04 to 5.3 ng/L/s in the linear mode. The analytical precision is 2.0 to 9.7% CV in the reflector mode, and 1.5 to 14.3% CV in the linear mode. A method comparison to a clinically employed LC-MS/MS assay for PRA determination showed excellent correlation within the linear range, with an R2 value of ≤0.98. This automated high throughput iMALDI platform has clinically suitable sensitivity, precision, linear range, and correlation with the standard method for PRA determination. Furthermore, the developed workflow based on the iMALDI technology can be used for the determination of other proteomic biomarkers. This article is part of a Special Issue entitled: Medical Proteomics.

Original languageEnglish
Pages (from-to)547-558
Number of pages12
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1854
Issue number6
DOIs
Publication statusPublished - 1 Jun 2015
Externally publishedYes

Keywords

  • Angiotensin-I
  • Automation
  • Hypertension
  • Immuno-MALDI (iMALDI)
  • Matrix assisted laser desorption/ionization (MALDI) mass spectrometry
  • Plasma renin activity (PRA)

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