Easy and Rapid Analysis of Protein Stability using the Prometheus NT.48

1 Feb 2016
Lois Manton-O'Byrne
Executive Editor

The Prometheus NT.48 is based on nanoDSF, an advanced Differential Scanning Fluorimetry technology, using intrinsic tryptophan and tyrosine fluorescence to measure protein stability with ultra-high resolution. The device uses capillaries for precise measurements, for easy sample handling, even for very viscous formulation conditions, and for providing a maintenance-free instrumentation.

Thermal or chemical stability can be investigated in a truly label-free setup by detecting changes in tryptophan or tyrosine fluorescence. NanoTemper’s dual UV-detector monitors the emission at 350 nm and 330 nm on-the-fly, and thus provides unmatched scanning speed and data point density. The ultra-high resolution data shows even minute differences in unfolding and is not affected by aggregation. As no secondary reporter fluorophores are required, protein solutions can be analyzed independent of buffer compositions, with a high dynamic protein concentration range from more than 250 mg/ml down to 5 µg/ml. This enables the analysis of detergent-solubilized membrane proteins as well as highly concentrated and viscous antibody formulations.

The Prometheus NT.48 can be optionally equipped with Backreflection Optics to detect aggregation of protein samples such as antibodies. The detection is based on an aggregation-induced light scattering. Owed to the high precision capillary format of the Prometheus series and automatic internal referencing, reproducibility and sensitivity of the aggregation detection are superior to conventional approaches. The combination of thermal stability detection and definition of aggregation onset temperatures allows for the most rapid, precise and information-rich analysis of biologicals, e.g. in formulation screenings or protein engineering projects.

Key Features of the Prometheus NT.48

  • Native DSF: no dye required
  • Tyrosine and tryptophan fluorescence
  • Dual UV system: 330 nm and 350 nm fluorescence is detected
  • 48 samples at a time
  • Ultra-high resolution: measure 48 capillaries in 3 seconds and see more unfolding transitions
  • Broad concentration range: from 5 µg/ml to more than 250 mg/ml
  • Temperature range: from 15 °C to 95 °C
  • Thermal and chemical denaturation
  • Maintenance-free instrument
  • Straightforward handling: simple sample preparation and intuitive software user interface

Key Features of the Backreflection Optics:

  • Optional backreflection optics: aggregation of protein samples is recorded
  • Ultra-high resolution: precisely determine aggregation onset temperatures
  • Broad concentration range: from 100 µg/ml to more than 100 mg/ml

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