Application Notes

Rapid quantification of unfolded proteins for quality control and optimization of storage conditions

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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

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