Boehringer Ingelheim Pharma – Michaela Blech

In order for proteins to manifest their proper biological and therapeutic effect, their conformational and structural integrity must be maintained at all stages of the development and commercialization process. Thus, for the development of Biotherapeutics it is essential to find optimal (solution) conditions to stabilize the Drug Product such that we can ensure its safety, biological function, structural integrity, and storage for long term under unfavourable and favourable conditions.

To meet the requirements of current clinical indications with respect to drug delivery, highly concentrated therapeutic protein drugs up to 100-200 mg/ml are necessary. In order to achieve such concentrations, a number of protein properties like solubility, self-association, solution viscosity, and protein aggregation have to be controlled. The large dynamic range of the Prometheus NT.48 allows for analyzing thermal unfolding and re-folding in solutions containing protein concentrations between 250 mg/ml down to few µg/ml. Moreover, measurements with rather viscous sample solutions as sometimes observed by highly concentrated formulations can be performed. Thus, it can be utilized for both, stability screening during early phases of drug discovery where only small amounts of protein are available, as well as for formulation screening campaigns of highly concentrated samples. The capabilities of the Prometheus NT.48 itself and in combination with other complementary biophysical tools enables for a better understanding of conformational and colloidal properties of the protein candidate and their impact on macroscopic solution properties. However, in some cases, the Prometheus NT.48 allowed us to perform thermal stability screening with otherwise structurally very complex proteins which were virtually not amenable to any orthogonal biophysical technique.

Moreover, the Prometheus instrument allows for label-free analysis of 48 samples simultaneously independent of their protein concentrations (high dynamic range) and selected solution conditions and/or –compositions. Unlike other techniques the Prometheus NT.48 measurements remained unaffected by any excipient, sugar, detergent or additive. Altogether, the Prometheus instrument enables for very flexible experimental design and provides maintenance-free instrumentation. In addition, our obtained data demonstrate very high reproducibility, consistency, the robustness and precision of this particular technology. The outstanding construction design allows for on-the-fly detection of fluorescence intensity resulting in impressive data point density that there is virtually no need for data fitting.

I personally also highly appreciate the very intuitive software allowing for rapidly setting up the experiment and evaluation of obtained data. The software being developed allows for rapid/easy data analysis, data interpretation and exportation to other data processing software.


 

Dr. Michaela Blech
Associate Director
Boehringer Ingelheim Pharma GmbH & Co. KG
Biberach, Germany