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Pitfalls in the determination of human acylated ghrelin plasma concentrations using a double antibody enzyme immunometric assay Trivedi, Arjun; Babic, Sandra; Chanoine, Jean-Pierre

Abstract

Objectives: To investigate the effect of hemolysis and protease inhibitors on acylated ghrelin (AG) concentrations measured with a double antibody enzyme immunometric assay that uses an acetylcholinesterase (AChE)-Fab’ conjugate. Design and Methods: Samples were hemolysed or treated with PHMB (phydroxymercuribenzoate), PMSF (phenylmethanesulfonylfluoride) or AEBSF (4-(2-Aminoethyl) benzenesulfonyl fluoride) to prevent AG degradation. Results: Hemolysis decreased AG concentrations. PHMB or PMSF did not affect the assay. The standard curve was abolished by AEBSF but rescued by addition of a washing step prior to the AChE-Fab' conjugate. Conclusions: Hemolysis and AEBSF may affect AG determination.

Item Metadata

Title
Pitfalls in the determination of human acylated ghrelin plasma concentrations using a double antibody enzyme immunometric assay
Creator
Trivedi, Arjun; Babic, Sandra; Chanoine, Jean-Pierre
Contributor
Child and Family Research Institute
Date Issued
2012-01
Description
Objectives: To investigate the effect of hemolysis and protease inhibitors on acylated ghrelin (AG) concentrations measured with a double antibody enzyme immunometric assay that uses an acetylcholinesterase (AChE)-Fab’ conjugate. Design and Methods: Samples were hemolysed or treated with PHMB (phydroxymercuribenzoate), PMSF (phenylmethanesulfonylfluoride) or AEBSF (4-(2-Aminoethyl) benzenesulfonyl fluoride) to prevent AG degradation. Results: Hemolysis decreased AG concentrations. PHMB or PMSF did not affect the assay. The standard curve was abolished by AEBSF but rescued by addition of a washing step prior to the AChE-Fab' conjugate. Conclusions: Hemolysis and AEBSF may affect AG determination.
Subject
Double antibody enzyme immunometric assay; Hemolysis; Protease Inhibitors; Acylated Ghrelin; Acetylcholinesterase
Genre
Article
Type
Text
Language
eng
Date Available
2015-10-24
Provider
Vancouver : University of British Columbia Library
Rights
Attribution-NonCommercial-NoDerivs 2.5 Canada
DOI
10.14288/1.0103472
URI
http://hdl.handle.net/2429/54878
Affiliation
Medicine, Faculty of; Pediatrics, Department of; Non UBC
Citation
Trivedi, A., Babic, S., & Chanoine, J. (2012). Pitfalls in the determination of human acylated ghrelin plasma concentrations using a double antibody enzyme immunometric assay. Clinical Biochemistry, 45(1–2), 178-180.
Publisher DOI
10.1016/j.clinbiochem.2011.10.023.
Peer Review Status
Reviewed
Scholarly Level
Faculty
Copyright Holder
Elsevier
Rights URI
http://creativecommons.org/licenses/by-nc-nd/2.5/ca/
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Attribution-NonCommercial-NoDerivs 2.5 Canada