Latent analysis of unmodified biomolecules and their complexes in solution with attomole detection sensitivity

Research output: Contribution to journalArticle

Abstract

The study of biomolecular interactions is central to an understanding of function, malfunction and therapeutic modulation of biological systems, yet often involves a compromise between sensitivity and accuracy. Many conventional analytical steps and the procedures required to facilitate sensitive detection, such as the incorporation of chemical labels, are prone to perturb the complexes under observation. Here we present a 'latent' analysis approach that uses chemical and microfluidic tools to reveal, through highly sensitive detection of a labelled system, the behaviour of the physiologically relevant unlabelled system. We implement this strategy in a native microfluidic diffusional sizing platform, allowing us to achieve detection sensitivity at the attomole level, determine the hydrodynamic radii of biomolecules that vary by over three orders of magnitude in molecular weight, and study heterogeneous mixtures. We illustrate these key advantages by characterizing a complex of an antibody domain in the solution phase and under physiologically relevant conditions.

Details

Authors
  • Emma V. Yates
  • Thomas Mueller
  • Luke Rajah
  • Erwin J. De Genst
  • Paolo Arosio
  • Sara Linse
  • Michele Vendruscolo
  • Christopher M. Dobson
  • Tuomas P. J. Knowles
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biochemistry and Molecular Biology
Original languageEnglish
Pages (from-to)802-809
JournalNature Chemistry
Volume7
Issue number10
Publication statusPublished - 2015
Publication categoryResearch
Peer-reviewedYes