Detection of Sotatercept (ACE‐011) in human serum by SAR‐PAGE and western single blotting
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| Deutscher übersetzter Titel: | Nachweis von Sotatercept (ACE-011) in Humanserum mittels SAR-PAGE und Western Single Blotting |
|---|---|
| Autor: | Reichel, Christian; Farmer, Letizia; Gmeiner, Günter; Walpurgis, Katja; Thevis, Mario |
| Erschienen in: | Drug testing and analysis |
| Veröffentlicht: | 10 (2018), 6, S. 927-937, Lit. |
| Format: | Literatur (SPOLIT) |
| Publikationstyp: | Zeitschriftenartikel |
| Medienart: | Elektronische Ressource (online) Gedruckte Ressource |
| Sprache: | Englisch |
| ISSN: | 1942-7603, 1942-7611 |
| DOI: | 10.1002/dta.2346 |
| Schlagworte: | |
| Online Zugang: | |
| Erfassungsnummer: | PU202011010135 |
| Quelle: | BISp |
Abstract des Autors
A method for the detection of Sotatercept (ACE‐011, ACVR2A‐Fc) in human serum is presented. The method is a modification of a recently published protocol for Luspatercept (ACE‐536, ACVR2B‐Fc), another erythropoiesis stimulating fusion protein. Out of 27 tested antibodies against either the extracellular domain of ACVR2A or the full‐length protein, only 4 antibodies bound strongly enough to Sotatercept for usage with immunoprecipitation followed by SAR‐PAGE and Western single blotting. The adapted protocol allows the detection of 0.1 ng/mL Sotatercept in just 50 μL human serum. None of the 3 commercial ACVR2‐ELISAs was able to detect Sotatercept and the 2 tested surrogate proteins, even in the μg/mL range. As for Luspatercept, only IPG‐IEF/2D‐PAGE generated discrete isoforms. Due to the long serum half‐life, the SAR‐PAGE method will be able to detect Sotatercept for several weeks and will be very useful in doping testing.