Universität zu Köln

Walpurgis, Katja, Thomas, Andreas ORCID: 0000-0003-1199-0743, Schaenzer, Wilhelm and Thevis, Mario (2016). Myostatin inhibitors in sports drug testing: Detection of myostatin-neutralizing antibodies in plasma/serum by affinity purification and Western blotting. Proteom. Clin. Appl., 10 (2). S. 195 - 206. WEINHEIM: WILEY-V C H VERLAG GMBH. ISSN 1862-8354

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Abstract

PurposeMyostatin is a key regulator of skeletal muscle growth and inhibition of its signaling pathway results in an increased muscle mass and function. The aim of this study was to develop a qualitative detection assay for myostatin-neutralizing antibodies for doping control purposes by using immunological approaches. Experimental designTo detect different types of myostatin-neutralizing antibodies irrespective of their amino acid sequence, an immunological assay specific for antibodies directed against myostatin and having a human Fc domain was established. Affinity purification and Western blotting strategies were combined to allow extracting and identifying relevant analytes from 200 L of plasma/serum in a non-targeted approach. The assay was characterized regarding specificity, linearity, precision, robustness, and recovery. ResultsThe assay was found to be highly specific, robust, and linear from 0.1 to 1 g/mL. The precision was successfully specified at three different concentrations and the recovery of the affinity purification was 58%. Conclusions and clinical relevanceWithin this study, an immunological detection assay for myostatin-neutralizing antibodies present in plasma/serum specimens was developed and successfully characterized. The presented approach can easily be modified to include other therapeutic antibodies and serves as proof-of-concept for the detection of antibody-based myostatin inhibitors in doping control samples.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Walpurgis, Katja UNSPECIFIED UNSPECIFIED UNSPECIFIED Thomas, Andreas UNSPECIFIED orcid.org/0000-0003-1199-0743 UNSPECIFIED Schaenzer, Wilhelm UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-285703
DOI:	10.1002/prca.201500043
Journal or Publication Title:	Proteom. Clin. Appl.
Volume:	10
Number:	2
Page Range:	S. 195 - 206
Date:	2016
Publisher:	WILEY-V C H VERLAG GMBH
Place of Publication:	WEINHEIM
ISSN:	1862-8354
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language SKELETAL-MUSCLE MASS; MUSCULAR-DYSTROPHY; THERAPEUTIC ANTIBODIES; IIB RECEPTOR; FIBER-TYPE; MICE; PROPEPTIDE; GROWTH; INCREASES; IDENTIFICATION Multiple languages Biochemical Research Methods; Biochemistry & Molecular Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/28570