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

Yates, Emma V.; Mueller, Thomas; Rajah, Luke; De Genst, Erwin J.; Arosio, Paolo; Linse, Sara LU ; Vendruscolo, Michele; Dobson, Christopher M. and Knowles, Tuomas P. J. (2015) In Nature Chemistry 7(10). p.802-809
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Nature Chemistry
volume
7
issue
10
pages
802 - 809
publisher
Nature Publishing Group
external identifiers
  • pmid:26391079
  • wos:000361753200009
  • scopus:84942314882
ISSN
1755-4330
DOI
10.1038/NCHEM.2344
language
English
LU publication?
yes
id
95bed059-abfb-475e-8bf3-8ccc9bf97f2e (old id 8058530)
date added to LUP
2015-10-21 08:49:06
date last changed
2017-11-05 03:00:43
@article{95bed059-abfb-475e-8bf3-8ccc9bf97f2e,
  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.},
  author       = {Yates, Emma V. and Mueller, Thomas and Rajah, Luke and De Genst, Erwin J. and Arosio, Paolo and Linse, Sara and Vendruscolo, Michele and Dobson, Christopher M. and Knowles, Tuomas P. J.},
  issn         = {1755-4330},
  language     = {eng},
  number       = {10},
  pages        = {802--809},
  publisher    = {Nature Publishing Group},
  series       = {Nature Chemistry},
  title        = {Latent analysis of unmodified biomolecules and their complexes in solution with attomole detection sensitivity},
  url          = {http://dx.doi.org/10.1038/NCHEM.2344},
  volume       = {7},
  year         = {2015},
}