Universität zu Köln

Reichel, Christian, Farmer, Letizia, Gmeiner, Guenter, Walpurgis, Katja and Thevis, Mario (2018). Detection of Sotatercept (ACE-011) in human serum by SAR-PAGE and western single blotting. Drug Test. Anal., 10 (6). S. 927 - 938. HOBOKEN: WILEY. ISSN 1942-7611

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Abstract

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 0.1ng/mL Sotatercept in just 50L human serum. None of the 3 commercial ACVR2-ELISAs was able to detect Sotatercept and the 2 tested surrogate proteins, even in the g/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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Reichel, Christian UNSPECIFIED UNSPECIFIED UNSPECIFIED Farmer, Letizia UNSPECIFIED UNSPECIFIED UNSPECIFIED Gmeiner, Guenter UNSPECIFIED UNSPECIFIED UNSPECIFIED Walpurgis, Katja UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-184715
DOI:	10.1002/dta.2346
Journal or Publication Title:	Drug Test. Anal.
Volume:	10
Number:	6
Page Range:	S. 927 - 938
Date:	2018
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1942-7611
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language LIGAND TRAP; POSTMENOPAUSAL WOMEN; ERYTHROPOIESIS; RECEPTOR; THERAPIES; ANEMIA; ACTRIIA-IGG1; ACE-536; AGENT Multiple languages Biochemical Research Methods; Chemistry, Analytical; Pharmacology & Pharmacy Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18471