Practical considerations regarding the choice of the best high-throughput assay

Carolin Mügge, Robert Kourist*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

All protein engineering studies include the stage of identifying and characterizing variants within a mutant library by employing a suitable assay or selection method. A large variety of different assay approaches for different enzymes have been developed in the last few decades, and the throughput performance of these assays vary considerably. Thus, the concept of a protein engineering study must be adapted to the available assay methods. This introductory review chapter describes different assay concepts on selected examples, including selection and screening approaches, detection of pH and cosubstrate changes, coupled enzyme assays, methods using surrogate substrates and selective derivatization. The given examples should guide and inspire the reader when choosing and developing own high-throughput screening approaches.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages189-208
Number of pages20
DOIs
Publication statusPublished - 1 Jan 2018

Publication series

NameMethods in Molecular Biology
Volume1685
ISSN (Print)1064-3745

Keywords

  • High-throughput screening
  • pH assay
  • Protein engineering
  • Selection
  • Selective derivatization
  • Surrogate substrates

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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