Arjun Trivedi1, Sandra Babic, Jean-Pierre Chanoine. 1. Department of Pediatrics, British Columbia Children's Hospital and Child and Family Research Institute, University of British Columbia, Vancouver, BC, Canada, V6H 3V4. atrivedi@interchange.ubc.ca
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 (p-hydroxymercuribenzoate), 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.
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 (p-hydroxymercuribenzoate), 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.
Authors: C Tezenas du Montcel; P Duriez; N Lebrun; D Grouselle; B de Grimaudet; R Dardennes; J Epelbaum; M Cuenca; O Viltart; P Gorwood; V Tolle Journal: Compr Psychoneuroendocrinol Date: 2022-05-20
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