Antibody-based strategies for the detection of Luspatercept (ACE-536) in human serum.
Drug Test Anal
; 9(11-12): 1721-1730, 2017 Nov.
Article
en En
| MEDLINE
| ID: mdl-28929587
ABSTRACT
Luspatercept (ACE-536, ACVR2B-Fc), a fusion protein consisting of the extracellular domain of ActRIIB receptor and the Fc-part of human immunoglobulin G1 (IgG1), is currently under clinical development (Phase III). It stimulates the formation of red blood cells and hence may be misused by athletes for doping purposes in the future. Several antibody-based strategies for the detection of Luspatercept and other ACVR2B-Fc fusion proteins in human serum were evaluated (ELISA; IEF-, SDS-, and SAR-PAGE followed by Western blotting; immunoprecipitation). Two methods led to useful results:
a commercial "soluble" ACTR-IIB ELISA, which also detected Luspatercept and other ACVR2B-Fc's, but showed no cross-reactivity with Sotatercept/ACVR2A-Fc's. The ELISA might be applied as fast screening tool (100 µL serum; limit of detection (LOD) ca 15.6 ng/mL). The second method uses a polyclonal ACVR2B-antibody for immunoprecipitation followed by SAR-PAGE and Western blotting with a monoclonal detection antibody (50 µL serum; LOD ca 1.0 ng/mL). It can be used for initial as well as for confirmatory testing. Due to the high doses (mg/kg) and long serum half-life of Luspatercept, both strategies may be useful in anti-doping control in the future. Copyright © 2017 John Wiley & Sons, Ltd.Palabras clave
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Proteínas Recombinantes de Fusión
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Fragmentos Fc de Inmunoglobulinas
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Activinas
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Receptores de Activinas Tipo II
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Anticuerpos Monoclonales
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Drug Test Anal
Asunto de la revista:
FARMACOLOGIA
Año:
2017
Tipo del documento:
Article