APPLICATION NOTE
Melanie Maschberger & Dennis Breitsprecher
NanoTemper Technologies GmbH
Munich, Germany
Abstract
The rigorous control of the quality of biological samples is of major importance in
pharmaceutical research. On the one hand, drug discovery projects with purified drug
targets such as kinases, receptors or integral membrane proteins require a consistent
quality of the proteins to ensure successful screening campaigns. On the other hand,
a consistent quality of biologicals such as antibodies or other therapeutic proteins is
essential in later stages of the drug development process, especially for tests in clinical
trials, and is finally required for approval of new drugs by federal agencies.
Here we demonstrate how the Prometheus NT.48 can be used to investigate the long-term
stability of proteins as well as to determine optimal handling conditions by quantifying
the fraction of unfolded proteins within seconds.
Keywords: Prometheus, protein stability, nanoDSF, stress testing, storage, % unfolded, IgG,
kinase, membrane protein
Introduction
The number of purified proteins that are used for diagnostic, academic and therapeutic
applications is constantly increasing. Especially the use of more challenging proteins such
as integral membrane proteins or conjugated antibodies is becoming more and more
widespread. Biophysical downstream analysis in drug discovery and in the development
of biologicals however is expensive, time consuming and o en requires a variety of
different instruments. Failure in those downstream or upstream processes due to an
insufficient protein quality can therefore have severe and expensive consequences [5].
Rapid quantification of unfolded proteins for quality
control and optimization of storage conditions