Comprehensive Comparison of the Capacity of Functionalized Sepharose, Magnetic Core, and Polystyrene Nanoparticles to Immuno-Precipitate Procalcitonin from Human Material for the Subsequent Quantification by LC-MS/MS
Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. The fast and accurate diagnosis of sepsis by procalcitonin (PCT) has emerged as an essential tool in clinical medicine. Although in use in the clinical laboratory for a long time, PCT quantification h...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-06-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/13/10963 |
| _version_ | 1797591483668234240 |
|---|---|
| author | Thomas Masetto Kai Matzenbach Thomas Reuschel Sebastian-Alexander Tölke Klaus Schneider Lea Marie Esser Marco Reinhart Laura Bindila Christoph Peter Matthias Grimmler |
| author_facet | Thomas Masetto Kai Matzenbach Thomas Reuschel Sebastian-Alexander Tölke Klaus Schneider Lea Marie Esser Marco Reinhart Laura Bindila Christoph Peter Matthias Grimmler |
| author_sort | Thomas Masetto |
| collection | DOAJ |
| description | Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. The fast and accurate diagnosis of sepsis by procalcitonin (PCT) has emerged as an essential tool in clinical medicine. Although in use in the clinical laboratory for a long time, PCT quantification has not yet been standardized. The International Federation of Clinical Chemistry working group on the standardization of PCT (IFCC-WG PCT) aims to provide an LC-MS/MS-based reference method as well as the highest metrological order reference material to address this diagnostic need. Here, we present the systematic evaluation of the efficiency of an immuno-enrichment method, based on functionalized Sepharose, magnetic-core, or polystyrene (latex) nano-particles, to quantitatively precipitate PCT from different human sample materials. This method may be utilized for both mass spectrometric and proteomic purposes. In summary, only magnetic-core nano-particles functionalized by polyclonal PCT antibodies can fulfil the necessary requirements of the international standardization of PCT. An optimized method proved significant benefits in quantitative and specific precipitation as well as in the subsequent LC-MS/MS detection of PCT in human serum samples or HeLa cell extract. Based on this finding, further attempts of the PCT standardization process will utilize a magnetic core-derived immuno-enrichment step, combined with subsequent quantitative LC-MS/MS detection. |
| first_indexed | 2024-03-11T01:39:03Z |
| format | Article |
| id | doaj.art-5e3c6f0a352e4fa5ba0d01cd23da91d4 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T01:39:03Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-5e3c6f0a352e4fa5ba0d01cd23da91d42023-11-18T16:46:05ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-06-0124131096310.3390/ijms241310963Comprehensive Comparison of the Capacity of Functionalized Sepharose, Magnetic Core, and Polystyrene Nanoparticles to Immuno-Precipitate Procalcitonin from Human Material for the Subsequent Quantification by LC-MS/MSThomas Masetto0Kai Matzenbach1Thomas Reuschel2Sebastian-Alexander Tölke3Klaus Schneider4Lea Marie Esser5Marco Reinhart6Laura Bindila7Christoph Peter8Matthias Grimmler9Institute of Molecular Medicine I, Medical Faculty, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, GermanyInstitute for Biomolecular Research, Hochschule Fresenius gGmbH, University of Applied Sciences, Limburger Straße 2, 65510 Idstein, GermanyInstitute for Biomolecular Research, Hochschule Fresenius gGmbH, University of Applied Sciences, Limburger Straße 2, 65510 Idstein, GermanyInstitute for Biomolecular Research, Hochschule Fresenius gGmbH, University of Applied Sciences, Limburger Straße 2, 65510 Idstein, GermanyInstitute for Biomolecular Research, Hochschule Fresenius gGmbH, University of Applied Sciences, Limburger Straße 2, 65510 Idstein, GermanyInstitute of Molecular Medicine I, Medical Faculty, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, GermanyGfA GmbH, Allgäuer Straße 1, 87459 Pfronten, GermanyClinical Lipidomics Unit, Institute of Physiological Chemistry, University Medical Center, 55131 Mainz, GermanyInstitute of Molecular Medicine I, Medical Faculty, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, GermanyDiaSys Diagnostic Systems GmbH, Alte Straße 9, 65558 Holzheim, GermanySepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. The fast and accurate diagnosis of sepsis by procalcitonin (PCT) has emerged as an essential tool in clinical medicine. Although in use in the clinical laboratory for a long time, PCT quantification has not yet been standardized. The International Federation of Clinical Chemistry working group on the standardization of PCT (IFCC-WG PCT) aims to provide an LC-MS/MS-based reference method as well as the highest metrological order reference material to address this diagnostic need. Here, we present the systematic evaluation of the efficiency of an immuno-enrichment method, based on functionalized Sepharose, magnetic-core, or polystyrene (latex) nano-particles, to quantitatively precipitate PCT from different human sample materials. This method may be utilized for both mass spectrometric and proteomic purposes. In summary, only magnetic-core nano-particles functionalized by polyclonal PCT antibodies can fulfil the necessary requirements of the international standardization of PCT. An optimized method proved significant benefits in quantitative and specific precipitation as well as in the subsequent LC-MS/MS detection of PCT in human serum samples or HeLa cell extract. Based on this finding, further attempts of the PCT standardization process will utilize a magnetic core-derived immuno-enrichment step, combined with subsequent quantitative LC-MS/MS detection.https://www.mdpi.com/1422-0067/24/13/10963sepsisprocalcitoninLC-MS/MS quantificationstandardizationimmuno-enrichmentbiopolymers |
| spellingShingle | Thomas Masetto Kai Matzenbach Thomas Reuschel Sebastian-Alexander Tölke Klaus Schneider Lea Marie Esser Marco Reinhart Laura Bindila Christoph Peter Matthias Grimmler Comprehensive Comparison of the Capacity of Functionalized Sepharose, Magnetic Core, and Polystyrene Nanoparticles to Immuno-Precipitate Procalcitonin from Human Material for the Subsequent Quantification by LC-MS/MS International Journal of Molecular Sciences sepsis procalcitonin LC-MS/MS quantification standardization immuno-enrichment biopolymers |
| title | Comprehensive Comparison of the Capacity of Functionalized Sepharose, Magnetic Core, and Polystyrene Nanoparticles to Immuno-Precipitate Procalcitonin from Human Material for the Subsequent Quantification by LC-MS/MS |
| title_full | Comprehensive Comparison of the Capacity of Functionalized Sepharose, Magnetic Core, and Polystyrene Nanoparticles to Immuno-Precipitate Procalcitonin from Human Material for the Subsequent Quantification by LC-MS/MS |
| title_fullStr | Comprehensive Comparison of the Capacity of Functionalized Sepharose, Magnetic Core, and Polystyrene Nanoparticles to Immuno-Precipitate Procalcitonin from Human Material for the Subsequent Quantification by LC-MS/MS |
| title_full_unstemmed | Comprehensive Comparison of the Capacity of Functionalized Sepharose, Magnetic Core, and Polystyrene Nanoparticles to Immuno-Precipitate Procalcitonin from Human Material for the Subsequent Quantification by LC-MS/MS |
| title_short | Comprehensive Comparison of the Capacity of Functionalized Sepharose, Magnetic Core, and Polystyrene Nanoparticles to Immuno-Precipitate Procalcitonin from Human Material for the Subsequent Quantification by LC-MS/MS |
| title_sort | comprehensive comparison of the capacity of functionalized sepharose magnetic core and polystyrene nanoparticles to immuno precipitate procalcitonin from human material for the subsequent quantification by lc ms ms |
| topic | sepsis procalcitonin LC-MS/MS quantification standardization immuno-enrichment biopolymers |
| url | https://www.mdpi.com/1422-0067/24/13/10963 |
| work_keys_str_mv | AT thomasmasetto comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms AT kaimatzenbach comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms AT thomasreuschel comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms AT sebastianalexandertolke comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms AT klausschneider comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms AT leamarieesser comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms AT marcoreinhart comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms AT laurabindila comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms AT christophpeter comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms AT matthiasgrimmler comprehensivecomparisonofthecapacityoffunctionalizedsepharosemagneticcoreandpolystyrenenanoparticlestoimmunoprecipitateprocalcitoninfromhumanmaterialforthesubsequentquantificationbylcmsms |