19F NMR as a Tool for Monitoring Individual Differentially Labeled Proteins in Complex Mixtures
Year: 2018
Type: article
Abstract: The ability to monitor the behavior of individual proteins in complex mixtures has many potential uses, ranging from analysis of protein interactions in highly concentrated solutions, modeling biological fluids or the intracellular environment, to optimizing biopharmaceutical co-formulations. Differential labeling NMR approaches, which traditionally use 15 N or 13 C isotope incorporation during recombinant expression, are not always practical in cases when endogenous proteins are obtained from an organism, or where the expression system does not allow for efficient labeling, especially for larger proteins. This study proposes differential labeling of proteins by covalent attachment of 19 F groups with distinct chemical shifts, giving each protein a unique spectral signature which can be monitored by 19 F NMR without signal overlap, even in complex mixtures, and without any interfering signals from the buffer or other unlabeled components. Parameters, such as signal intensities, translational diffusion coefficients, and transverse relaxation rates, which report on the behavior of individual proteins in the mixture, can be recorded even for proteins as large as antibodies at a wide range of concentrations.
Source: Molecular Pharmaceutics
Authors: John M. Edwards, Jeremy P. Derrick, Christopher F. van der Walle, Alexander P. Golovanov
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: green
Funder: MedImmune
OpenAlex ID: W2805197574