Prometheus Panta for biologics stability profiling with particle sizing and molecular weight determination by NanoTemper Technologies

Features

• Monitor the stability profile of a wide range of protein-based biologic candidates, including monoclonal antibodies (mAbs), their derivatives (e.g. Fabs), enzymes, or receptor blockers
• Domain-specific information about your candidate molecule when you measure thermal unfolding, particle sizing, self-association, and aggregation simultaneously throughout the entire thermal ramp
• High-resolution data that reveals even subtle differences in candidate behavior, exposing potential liabilities in your drug candidates
• Flexible throughput and a few choices for sample handling

Parameters that Prometheus Panta measures:

Dynamic Light Scattering (DLS)

SIZE ANALYSIS

T_size (from growth of r_H in cumulants fit): Temperature at which average candidate size begins to increase, indicating the beginning of the unfolding process

Average scattering intensity: Identifies if sample concentration is too high or low for proper size distribution analysis

CUMULANT OR SIZE DISTRIBUTION/REGULARIZATION ANALYSIS

r_H: Hydrodynamic radius shows the size of your candidate in its solvated state

PDI: Polydispersity index, represents distribution of size populations and indicates if a sample preparation is pure and contains well-folded candidate proteins/mAbs

SELF-INTERACTION ANALYSES

k_D: Self-interaction parameter, used to predict the propensity of a biologic candidate to aggregate once it is at concentrations required for clinical administration

D₀ (*derived from k_D analysis): Theoretical diffusion constant at concentration = 0, a ranking parameter for finding optimal formulation conditions for a candidate

nanoDSF

THERMAL UNFOLDING

T_m (for 330 nm, 350 nm, and ratio): Melting temperature, or point at which 50% of your protein is unfolded; higher T_m values indicate greater thermal stability

T_onset/T_on (for ratio): Temperature at which unfolding begins; higher T_on values indicate increased thermal stability, and it is considered favorable to have a sharper slope at the unfolding inflection point, which occurs when the T_on is close in value to the T_m

E_a (*derived from thermal unfolding data): Activation energy of unfolding, a parameter that informs on the stability of a protein in a given buffer environment

Reversibility of unfolding: Point at which aggregated candidate protein cannot be recovered and refolded without other chemical means

Backreflection

AGGREGATION

T_turbidity: Onset temperature of amorphous aggregate (>12.5 nm radius) formation

SLS

SIZING INFORMATION

Molecular weight: Average molecular weight of all particles in solution

SELF-ASSOCIATION AND AGGREGATION PROPENSITY

T_scattering: Temperature at which scattering intensity begins to change, indicating increased size and aggregation

B₂₂: Second virial coefficient, used to extrapolate self-association behavior at higher concentrations such as those used for subcutaneous administration