Combination of high throughput and structural screening to assess protein stability - A screening perspective

Christin Pohl*, Sujata Mahapatra, Alina Kulakova, Werner Streicher, Günther H.J. Peters, Allan Nørgaard, Pernille Harris*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

High throughput screening to measure the stability of industrially relevant proteins and their variants is necessary for quality assessment in the development process. Advances in automation, measurement time and sample consumption for many techniques allow rapid measurements with minimal amount of protein. However, many methods include automated data analysis, potentially neglecting important aspects of the proteińs behavior in certain conditions. In this study we implement small angle X-ray scattering (SAXS), typically not used to assess protein behavior in industrial screening, in a high throughput screening workflow to address problems of contradicting results and reproducibility among different high throughput methods. As a case study we use the lipases of Thermomyces lanuginosus and Rhizomucor miehei, widely used industrial biocatalysts. We show that even the initial analysis of the SAXS data without performing any time-consuming modelling provide valuable information on interparticle interactions. We conclude that recent advances in automation and data processing, have enabled SAXS to be used more widely as a tool to gain in-depth knowledge highly useful for protein formulation development. This is especially relevant in light of increasing accessibility to SAXS due to the commercial availability of benchtop instruments.
Original languageEnglish
JournalEuropean Journal of Pharmaceutics and Biopharmaceutics
Volume171
Number of pages10
ISSN0939-6411
DOIs
Publication statusPublished - 2022

Keywords

  • High throughput screening
  • Drug screening
  • Small-angle X-ray scattering (SAXS)
  • Protein stability
  • Protein-protein interaction
  • Protein aggregation
  • Protein engineering

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