Development of Broadly Applicable Reference EVs Expressing Horseradish Peroxidase
ABSTRACT Extracellular vesicles (EVs) hold great promise as circulating biomarkers for diagnostic and therapeutic approaches. Thus, many research groups world‐wide investigate important aspects of EVs including their biology and medical significance. For this, a large number of procedures and protoc...
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| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2025-08-01
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| Series: | Journal of Extracellular Vesicles |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jev2.70115 |
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| _version_ | 1849222181707317248 |
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| author | Daniela Waas Marc Juraschitz Yen‐Fu Adam Chen Harald Waltenberger Wolfgang Hammerschmidt Reinhard Zeidler Sonja Molinaro Kathrin Gärtner |
| author_facet | Daniela Waas Marc Juraschitz Yen‐Fu Adam Chen Harald Waltenberger Wolfgang Hammerschmidt Reinhard Zeidler Sonja Molinaro Kathrin Gärtner |
| author_sort | Daniela Waas |
| collection | DOAJ |
| description | ABSTRACT Extracellular vesicles (EVs) hold great promise as circulating biomarkers for diagnostic and therapeutic approaches. Thus, many research groups world‐wide investigate important aspects of EVs including their biology and medical significance. For this, a large number of procedures and protocols has been established making it difficult to almost impossible to compare and replicate results. Consequently, diagnostic tests remain problematic to interpret, mainly because the use of reliable reference EVs as a qualified standard has not yet gained widespread acceptance. Beyond doubt, such reference EVs are key to assess EV preparations quantitatively and to establish robust quality control processes to ensure overall quality and validity of data. To further advance the establishment of such controls, we designed and generated a new class of reference EVs expressing horseradish peroxidase (HRP) to facilitate simple and reliable EV tracing during isolation and standardization of EV purification and downstream analysis processes. HRP+ EVs can be quantified easily and with unmatched sensitivity either directly via measuring HRP activity or indirectly via immunodetection of HRP on the EV surface. We demonstrate that HRP+ EVs allow the reliable quantification of absolute EV numbers in biological or medical samples to normalize clinical specimens in liquid biopsies. |
| format | Article |
| id | doaj-art-e11eceb87fe24d92ab9aeeae3e55e799 |
| institution | Kabale University |
| issn | 2001-3078 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Vesicles |
| spelling | doaj-art-e11eceb87fe24d92ab9aeeae3e55e7992025-08-26T07:24:44ZengWileyJournal of Extracellular Vesicles2001-30782025-08-01148n/an/a10.1002/jev2.70115Development of Broadly Applicable Reference EVs Expressing Horseradish PeroxidaseDaniela Waas0Marc Juraschitz1Yen‐Fu Adam Chen2Harald Waltenberger3Wolfgang Hammerschmidt4Reinhard Zeidler5Sonja Molinaro6Kathrin Gärtner7Eximmium Biotechnologies GmbH Munich GermanyMicrocoat Biotechnologie GmbH Bernried GermanyEximmium Biotechnologies GmbH Munich GermanyMicrocoat Biotechnologie GmbH Bernried GermanyResearch Unit Gene Vectors, Helmholtz Munich German Research Center for Environmental Health Munich GermanyInstitute of Structural Biology, Helmholtz Munich German Research Center for Environmental Health Munich GermanyMicrocoat Biotechnologie GmbH Bernried GermanyEximmium Biotechnologies GmbH Munich GermanyABSTRACT Extracellular vesicles (EVs) hold great promise as circulating biomarkers for diagnostic and therapeutic approaches. Thus, many research groups world‐wide investigate important aspects of EVs including their biology and medical significance. For this, a large number of procedures and protocols has been established making it difficult to almost impossible to compare and replicate results. Consequently, diagnostic tests remain problematic to interpret, mainly because the use of reliable reference EVs as a qualified standard has not yet gained widespread acceptance. Beyond doubt, such reference EVs are key to assess EV preparations quantitatively and to establish robust quality control processes to ensure overall quality and validity of data. To further advance the establishment of such controls, we designed and generated a new class of reference EVs expressing horseradish peroxidase (HRP) to facilitate simple and reliable EV tracing during isolation and standardization of EV purification and downstream analysis processes. HRP+ EVs can be quantified easily and with unmatched sensitivity either directly via measuring HRP activity or indirectly via immunodetection of HRP on the EV surface. We demonstrate that HRP+ EVs allow the reliable quantification of absolute EV numbers in biological or medical samples to normalize clinical specimens in liquid biopsies.https://doi.org/10.1002/jev2.70115extracellular vesicleshorseradish peroxidasenormalizationquality controlstandardization |
| spellingShingle | Daniela Waas Marc Juraschitz Yen‐Fu Adam Chen Harald Waltenberger Wolfgang Hammerschmidt Reinhard Zeidler Sonja Molinaro Kathrin Gärtner Development of Broadly Applicable Reference EVs Expressing Horseradish Peroxidase Journal of Extracellular Vesicles extracellular vesicles horseradish peroxidase normalization quality control standardization |
| title | Development of Broadly Applicable Reference EVs Expressing Horseradish Peroxidase |
| title_full | Development of Broadly Applicable Reference EVs Expressing Horseradish Peroxidase |
| title_fullStr | Development of Broadly Applicable Reference EVs Expressing Horseradish Peroxidase |
| title_full_unstemmed | Development of Broadly Applicable Reference EVs Expressing Horseradish Peroxidase |
| title_short | Development of Broadly Applicable Reference EVs Expressing Horseradish Peroxidase |
| title_sort | development of broadly applicable reference evs expressing horseradish peroxidase |
| topic | extracellular vesicles horseradish peroxidase normalization quality control standardization |
| url | https://doi.org/10.1002/jev2.70115 |
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