Chapter 5. Continuous-Flow Systems for Ligand Binding and Enzyme Inhibition Assays Based on Mass Spectrometry

  1. Prof. Dr. Klaus T. Wanner and
  2. Dr. Georg Höfner
  1. Hubertus Irth

Published Online: 24 MAY 2007

DOI: 10.1002/9783527610907.ch5

Mass Spectrometry in Medicinal Chemistry: Applications in Drug Discovery

Mass Spectrometry in Medicinal Chemistry: Applications in Drug Discovery

How to Cite

Irth, H. (2007) Continuous-Flow Systems for Ligand Binding and Enzyme Inhibition Assays Based on Mass Spectrometry, in Mass Spectrometry in Medicinal Chemistry: Applications in Drug Discovery (eds K. T. Wanner and G. Höfner), Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany. doi: 10.1002/9783527610907.ch5

Editor Information

  1. Ludwig-Maximilians-University Munich, Department of Pharmacy, Center of Drug Research, Butenandtstr., 81377 Munich, Germany

Author Information

  1. Department of Analytical Chemistry & Applied Spectroscopy, Vrije Universiteit Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands

Publication History

  1. Published Online: 24 MAY 2007
  2. Published Print: 23 FEB 2007

Book Series:

  1. Methods and Principles in Medicinal Chemistry

Book Series Editors:

  1. Prof. Dr. Raimund Mannhold3,
  2. Prof. Dr. Hugo Kubinyi4 and
  3. Prof. Dr. Gerd Folkers5

Series Editor Information

  1. 3

    Molecular Drug Research Group, Heinrich-Heine-Universität, Universitätsstrasse 1, 40225 Düsseldorf, Germany

  2. 4

    Donnersbergerstrasse 9, 67256 Weisenheim am Sand, Germany

  3. 5

    Collegium Helveticum, STW/ETH Zurich, 8092 Zurich, Switzerland

ISBN Information

Print ISBN: 9783527314560

Online ISBN: 9783527610907

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Keywords:

  • mass spectrometry;
  • continuous-flow enzyme assays based on mass spectrometry;
  • assay principles;
  • ESI-MS assay of cathepsin B;
  • ESI-MS assay of acetylcholinesterase;
  • miniaturization of electrospray MS assays;
  • continuous-flow ligand binding assays based on mass spectrometry;
  • optimization of MS conditions;
  • monitoring bioactive compounds;
  • continuous-flow multi-protein binding assays using electrospray MS

Summary

This chapter contains sections titled:

  • Introduction

  • Continuous-flow Enzyme Assays Based on Mass Spectrometry

    • Assay Principle

    • ESI-MS Assay of Cathepsin B

      • MS Assay Development for Cathepsin B

      • Compatibility of Cathepsin B Assay with MS Detection

      • On-line Coupling of MS-based Cathepsin B Assay to HPLC

      • Screening of Natural Products for Cathepsin B Activity

    • ESI-MS Assay of Acetylcholinesterase

      • MS Assay Development for Acetylcholinesterase

      • Assay Validation and Stability

      • Screening of Natural Products for Acetylcholinesterase Activity

    • Miniaturization of Electrospray MS Assays

      • Chip-based Electrospray MS Assays

      • Chip Performance

      • Sensitivity of the Chip-based MS Screening System

  • Continuous-flow Ligand Binding Assays Based on Mass Spectrometry

    • Assay Principle

    • Optimization of MS Conditions

    • On-line Continuous-flow Biochemical Interaction

    • Monitoring Bioactive Compounds

    • Antibody–Antigen Interactions

    • Continuous-flow Multi-protein Binding Assays Using Electrospray MS

  • MS Assay Based on Dissociation of Isolated Protein–Ligand Complexes

    • Assay Set-up

    • Flow Injection Label-free MS Assay

    • Flow Injection Label-free MS Assay Screening of Natural Extracts

  • Future Prospects