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:
CreatorsEmailORCIDORCID Put Code
Walpurgis, KatjaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Thomas, AndreasUNSPECIFIEDorcid.org/0000-0003-1199-0743UNSPECIFIED
Schaenzer, WilhelmUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Thevis, MarioUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
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:
KeywordsLanguage
SKELETAL-MUSCLE MASS; MUSCULAR-DYSTROPHY; THERAPEUTIC ANTIBODIES; IIB RECEPTOR; FIBER-TYPE; MICE; PROPEPTIDE; GROWTH; INCREASES; IDENTIFICATIONMultiple languages
Biochemical Research Methods; Biochemistry & Molecular BiologyMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/28570

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