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Strategy for peptide quantification using LC-MS in regulated bioanalysis: case study with a glucose-responsive

Yang Xu1, Dina Goykhman1, Ming Wang1

  • 1Pharmacokinetics, Pharmacodynamics & Drug Metabolism, Regulated Bioanalysis, Merck & Co., Inc., 770 Sumneytown Pike, WP75B-300, West Point, PA 19486, USA.

Bioanalysis
|August 1, 2018
PubMed
Summary

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) offers advanced peptide quantification but faces challenges. This study presents a validated LC-MS/MS strategy for hybrid assays, exemplified by glucose-responsive insulin (MK-2640).

Keywords:
GRIglucose-responsive insulinhybrid assaypeptide quantification

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Area of Science:

  • Analytical Chemistry
  • Biotechnology
  • Pharmacology

Background:

  • Traditional ligand-binding assays are being replaced by LC-MS/MS for peptide quantification.
  • LC-MS/MS presents challenges in sensitivity, specificity, and ruggedness, particularly for hybrid assay formats.
  • Regulatory guidance for LC-MS/MS hybrid assays is lacking.

Purpose of the Study:

  • To present a robust strategy for LC-MS/MS method development and assay validation.
  • To address the challenges associated with hybrid assay formats in peptide quantification.
  • To provide a case study using MK-2640, a glucose-responsive insulin, in various matrices.

Main Methods:

  • Method development and assay validation strategy for LC-MS/MS.
  • Application of immunoaffinity purification for rat/dog GLP studies.
  • Solid-phase extraction (SPE) for clinical studies.
  • Quantification of intact MK-2640.

Main Results:

  • Successful development and validation of an LC-MS/MS method for MK-2640.
  • Demonstrated applicability of the strategy across multiple biological matrices.
  • Immunoaffinity purification and SPE effectively supported different study types.

Conclusions:

  • The presented strategy provides a framework for developing and validating LC-MS/MS assays for peptide quantification.
  • The approach is suitable for hybrid assay formats and complex matrices.
  • This work aids in navigating regulatory challenges for advanced peptide analysis.