Guide: Prometheus Panta Parameters Explained

Multiplexing Optical Methods in Prometheus™ Panta for Comprehensive Protein Characterization

When it comes to understanding protein behavior, one technique is rarely enough. Proteins are complex, and their stability, purity, size, and aggregation tendencies all play critical roles in research and development. That’s why the Prometheus™ Panta from NanoTemper stands out — by combining multiple optical methods into a single platform, it enables a truly comprehensive characterization of proteins.

Why Multiplexing Optical Methods?
Protein characterization demands more than just one type of measurement. By multiplexing optical techniques such as nanoDSF, DLS, SLS, and backreflection, Prometheus Panta provides a multidimensional profile of your protein samples. This allows researchers to make more informed and confident decisions when selecting candidates for further development, troubleshooting stability issues, or optimizing formulations.

Let’s break down the key parameters measured by each method and why they matter.

Your Guide: Prometheus Panta Parameters Explained

Below, we provide a comprehensive overview of the key analytical parameters, definitions and practical relevance for protein characterization, with links to dedicated deep dive articles.

Use this guide to better interpret your results and make confident, data-driven decisions throughout your protein analysis workflows.

Conformational Stability: nanoDSF
Prometheus Panta uses nanoDSF (differential scanning fluorimetry) to assess thermal and chemical unfolding of proteins. Important parameters include:

• T_m (melting temperature): The point at which 50% of a protein or domain is unfolded. This is critical for ranking candidates based on thermostability. 
• T_onset: The temperature at which unfolding begins. A higher Tonset generally indicates greater protein stability.
• Ea (Activation energy): Reflects how easily a protein unfolds. A higher Ea suggests slower unfolding, important for predicting long-term stability.
• Initial ratio (350nm/330nm fluorescence): Indicates the initial fold state, giving insight into the percentage of folded protein.
• C_m or C₅₀: Concentration of denaturant at which 50% unfolding occurs, useful for chemical stability.
• ΔG (Gibbs free energy): Shows the thermodynamic stability by comparing folded vs. unfolded states.

Purity: DLS
Dynamic Light Scattering (DLS) in Prometheus Panta helps determine the Polydispersity Index (PDI). This metric reveals the heterogeneity of sample populations — critical for spotting large aggregates or multiple protein populations.

Sizing: DLS & SLS
Prometheus Panta uses DLS and Static Light Scattering (SLS) to explore size-related parameters:

• r_H (Hydrodynamic radius): Derived from diffusion coefficients, it provides an average particle size.
• T_size (Temperature where size increases): Indicates when particles begin to unfold, oligomerize, or aggregate.
• MW (Molecular weight): Offers insights into whether your sample contains monomers, oligomers, or aggregates, helping detect modifications or stress-induced changes.

Aggregation & Self-Association: SLS & DLS
Monitoring aggregation is vital to ensure therapeutic viability. Prometheus Panta examines:

• T_turbidity: Temperature at which turbidity (large aggregates) begins to increase.
• T_scattering: Temperature at which scattering (even small aggregates) rises, offering sensitivity beyond turbidity.
• B₂₂ (Self-interaction parameter): Indicates attractive or repulsive interactions. Negative B22 suggests a higher aggregation risk.
• kD (Diffusion interaction parameter): Assesses weak interactions. Negative values often point to aggregation tendencies at high concentrations.

For more resources and to download this guide as a PDF, visit our support center.

Efficiency in Protein Characterization
The power of Prometheus Panta lies in its ability to consolidate all these measurements into a single workflow. This means less sample consumption, more data per experiment, and a holistic understanding of your protein’s properties.

Whether you’re screening candidates, optimizing formulations, or ensuring stability, Multiplexing Optical Methods with Prometheus Panta accelerates your path to success.

Explore how Prometheus™ Panta can streamline your protein characterization pipeline and deliver richer insights for your research.