APPLICATION NOTE
Dennis Breitsprecher
1
, Pawel Linke
1
, Anna Schulze
1
, Franziska Söltl
1
,
Patrick Garidel
2
, Michaela Blech
2
1
NanoTemper Technologies GmbH, Munich, Germany
2
Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany
Abstract
One of the most important parameters in the development of therapeutic biologics is
their long-term stability. While a er purification being seemingly stable in a variety of
formulations, many antibodies display very slow aggregation kinetics over time. This
gradual aggregation could thus far only be evaluated by monitoring monomer contents
and aggregates over months or even years. Predictive methods are therefore urgently
needed to speed up the development of biologics.
Here we demonstrate that the Prometheus NT.Plex by NanoTemper Technologies can
be used for predicting long-term stability in only a few hours. The approach uses a
combination of thermal and chemical unfolding analysis in a high-throughput setting.
We show that chemical denaturation is a tool which can determine folding enthalpies
of monoclonal antibodies (mAbs) to predict their long-term stability in formulation
screenings. Using the Prometheus NT.Plex nanoDSF instrument with aggregation
detection optics, we screened 5 formulations at different mAb concentrations for their
thermal and chemical stability. The obtained unfolding data correlates with long-term
turbidity and monomer content over time, showing that the Prometheus NT.Plex can be
used to rapidly predict the long-term stability of biologics within 1 day.
Keywords: protein stability, aggregation propensity prediction, IgG, formulation, biologics,
thermal and chemical stability, ∆G
Getting the full picture: predicting the aggregation
propensity of mAbs using chemical and thermal
denaturation on a single, fully automated platform