Quantification of serum apolipoproteins A-I and B-100 in clinical samples using an automated SISCAPA-MALDI-TOF-MS workflow

Methods : a Companion to Methods in Enzymology
Irene van den BroekChrista M Cobbaert

Abstract

A fully automated workflow was developed and validated for simultaneous quantification of the cardiovascular disease risk markers apolipoproteins A-I (apoA-I) and B-100 (apoB-100) in clinical sera. By coupling of stable-isotope standards and capture by anti-peptide antibodies (SISCAPA) for enrichment of proteotypic peptides from serum digests to matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS detection, the standardized platform enabled rapid, liquid chromatography-free quantification at a relatively high throughput of 96 samples in 12h. The average imprecision in normo- and triglyceridemic serum pools was 3.8% for apoA-I and 4.2% for apoB-100 (4 replicates over 5 days). If stored properly, the MALDI target containing enriched apoA-1 and apoB-100 peptides could be re-analyzed without any effect on bias or imprecision for at least 7 days after initial analysis. Validation of the workflow revealed excellent linearity for daily calibration with external, serum-based calibrators (R(2) of 0.984 for apoA-I and 0.976 for apoB-100 as average over five days), and absence of matrix effects or interference from triglycerides, protein content, hemolysates, or bilirubins. Quantification of apoA-I in 93 normo- and h...Continue Reading

References

May 5, 2009·Molecular & Cellular Proteomics : MCP·Michael A KuzykChristoph H Borchers
Jun 23, 2009·Journal of Immunological Methods·Andrew N Hoofnagle, Mark H Wener
Dec 1, 2007·Proteomics. Clinical Applications·Jian JiangChristoph H Borchers
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Aug 24, 2013·Proteomics. Clinical Applications·Uta CeglarekJoachim Thiery
Dec 17, 2014·Journal of Proteome Research·Irene van den BroekChrista M Cobbaert

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Citations

Jan 2, 2016·Journal of Proteomics·Pascal SteffenHartmut Schlüter
Aug 22, 2015·Bioanalysis·Irene van den Broek, William D van Dongen
Dec 19, 2015·Proteomics. Clinical Applications·Cédric MesminBruno Domon
Jun 5, 2015·Methods : a Companion to Methods in Enzymology·Benoit Coulombe, Marie-Soleil Gauthier
Nov 14, 2015·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Tove BoströmSophia Hober
Oct 25, 2016·Journal of Proteomics·Christopher A TothJohn R Barr
May 20, 2020·Expert Review of Proteomics·Alain Van GoolMarei Sammar
Sep 6, 2017·Expert Review of Proteomics·Irene van den BroekJennifer E Van Eyk
Nov 2, 2016·Expert Review of Proteomics·L Renee RuhaakChrista M Cobbaert
Nov 9, 2021·Mass Spectrometry Reviews·Erica GianazzaCristina Banfi

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