Comparison of Targeted Mass Spectrometry Techniques with an Immunoassay: A Case Study for HSP90α
| Autor: | Theo M Luidert, Lennard J. M. Dekker, Coşkun Güzel, Rainer Bischoff, Alexander P. Boichenko, Ate G.J. van der Zee, Natalia Govorukhina, Christoph Stingl |
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| Přispěvatelé: | Analytical Biochemistry, Targeted Gynaecologic Oncology (TARGON), Medicinal Chemistry and Bioanalysis (MCB), Neurology |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
musculoskeletal diseases Adult Proteomics PROTEINS viruses Clinical Biochemistry PRM Mass spectrometry Tandem mass spectrometry 03 medical and health sciences Limit of Detection Tandem Mass Spectrometry medicine Journal Article HSP90 Humans ASSAYS QUANTITATIVE PROTEOMICS Amino Acid Sequence HSP90 Heat-Shock Proteins Detection limit Immunoassay Reproducibility Chromatography medicine.diagnostic_test PLASMA Chemistry virus diseases Reproducibility of Results Repeatability QUANTIFICATION CANCER humanities Peptide Fragments Triple quadrupole mass spectrometer 030104 developmental biology Targeted mass spectrometry CELLS Proteolysis Female human activities HIGH-RESOLUTION Chromatography Liquid |
| Zdroj: | Proteomics. Clinical Applications, 12(1):1700107. WILEY-V C H VERLAG GMBH Proteomics Clinical Applications, 12(1):1700107. Wiley-VCH |
| ISSN: | 1862-8354 1862-8346 |
| Popis: | PURPOSE: The objective of this study was to better understand factors governing the variability and sensitivity in SRM and PRM, compared to immunoassay.EXPERIMENTAL DESIGN: A 2D-LC-MS/MS-based SRM and PRM assay was developed for quantitative measurements of HSP90α in serum. Forty-three control sera were compared by SRM, PRM and ELISA following the manufacturer's instructions. Serum samples were trypsin-digested and fractionated by SCX chromatography prior to SRM and PRM measurements. Analytical parameters such as linearity, LOD, LOQ, repeatability and reproducibility of the SRM, PRM and ELISA were determined.RESULTS: PRM data obtained by high-resolution mass spectrometry correlated better with ELISA measurements than SRM data measured on a triple quadrupole mass spectrometer. While all three methods (SRM, PRM, ELISA) were able to quantify HSP90α in serum at the ng/mL level, the use of PRM on a high-resolution mass spectrometer reduced variation and showed comparable sensitivity to immunoassay.CONCLUSIONS AND CLINICAL RELEVANCE: Using fractionation, it is possible to measure ng/mL levels of HSP90α in a reproducible, selective and sensitive way using PRM in serum. This opens up the possibility to use PRM in a multiplexed way as an attractive alternative for immunoassays without the use of antibodies or comparable binders. This article is protected by copyright. All rights reserved. |
| Databáze: | OpenAIRE |
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