Strategy for peptide quantification using LC–MS in regulated bioanalysis: case study with a glucose-responsive insulin. (31st July 2018)

• Record Type: Journal Article
• Authors: Xu, Yang; Goykhman, Dina; Wang, Ming; Jackson, Tonya; Xie, Iris; Willson, Kenneth; Breidinger, Sheila; Schuck, Helengrace; Harrelson, Jane; Woolf, Eric J
• Is Part Of: Bioanalysis. Volume 10:Number 15(2018)
• Page Start: 1207
• Page End: 1220
• Publication Date: 2018-07-31
• Subjects: glucose-responsive insulin -- GRI -- hybrid assay -- peptide quantification; Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio-2018-0089
• Languages: English
• ISSNs: 1757-6180

Abstract

Aim: Advances in technology have led to a shift for peptide quantification from traditional ligand-binding assays to LC–MS/MS-based analysis, which presents challenges, in other assay sensitivity, specificity and ruggedness, in addition to lacking of regulatory guidance, especially for the hybrid assay format.Methodology & results: This report communicates a strategy that has been employed in our laboratories for method development and assay validation, and exemplified in a case study of MK-2640, a glucose-responsive insulin, in multiple matrices. Intact MK-2640 was monitored, while immunoaffinity purification and SPE were used to support the rat/dog GLP and clinical studies, respectively. The rationale and considerations behind our approach, as well as the acceptance criteria applied to the assay validation are discussed.