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NanoTemper Prometheus

The new gold standard for precise characterization of protein stability

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Prometheus has been chosen by the scientific community because it consistently delivers high-resolution data that reveals liabilities in drug candidates missed by other technologies. No other system offers simultaneous, multi-parameter characterization throughout the entire run — measuring thermal unfolding, particle sizing, and aggregation — as well as chemical denaturation for a stability profile of unmatched detail.


Prometheus consistently delivers trustworthy, high-resolution stability characterization

Get precise, high-resolution data and reliable results every time

Measuring with precision matters when there are tiny differences between your candidates. Prometheus delivers the kind of stability data you need to clearly see those differences and makes it easier to decide which candidates are best to work with moving forward.


Perform simultaneous measurements with the same samples throughout the entire run

Since you’re doing multi-parameter characterization on the same samples in the same run, you gain a deeper understanding of your molecule’s behavior when correlating your conformational and colloidal stability results.


Choose your throughput, and then choose manual or automated

Be prepared to handle any project that comes your way. Characterizing stability at different checkpoints in your workflow calls for flexible throughput — early development stages require characterizing more candidates than later, during formulation. Choose to run any number of samples from 1 to 48 or 24 at a time. Add automation and do 1536 before having to reload.


Measure label-free under native conditions

Prometheus detects intrinsic fluorescence so you don’t have to introduce dyes and risk interference. It gets better — no need for sample dilution or special buffer conditions means very little or no sample prep. And, have viscous samples? No problem.



Discover how Prometheus’s specificity, sensitivity, accuracy, precision, and limit of detection makes a difference in biologics stability characterization

Predict biologics developability profiles through early-stage screening.

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Perform comparability studies to validate that changes to manufacturing processes or sites don’t affect the drug product.

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Facilitate and validate engineered stability enhancements. Correlate these with optimization the biologic’s behavior in storage and accelerated degradation assays.

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Where Prometheus is making a difference?


Drug discovery & development

Use every time stability needs to be characterized and monitored to rank molecule candidates


Structural biology

Maximize protein solubility and stability before crystallization to dramatically increase its probability of crystallization


Gene Therapy

Differentiate viral vectors serotypes during development and manufacturing based on their thermal stability profiles

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Biologics discovery & development

  •   - Evaluate target-ligand complexes before affinity-based screening

  •   - Developability assessment

  •   - Pre-formulation and formulation

  •   - Production & downstream process development

  •   - Comparability studies

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Protein expression & purification

Monitor the stability of recombinant proteins during selection of expression clones and chromatographic isolation and purification



Thermal shift assays

Screen compounds during drug development for shifts in thermal unfolding to eliminate compounds that decrease stability



Monitor stability of recombinant proteins during chromatographic isolation and purification



A system for every one of your needs, present or future

It’s important to take into account your evolving needs for stability characterization and throughput. Choosing fully-featured Prometheus Panta gives you the peace of mind knowing you’ll be ready for projects that demand simultaneous, multi-parameter characterization. If having a path to automation is what you and your stakeholders have in mind, go with the Prometheus NT.Plex.


♦ Prometheus Panta

Choose Prometheus Panta if you want everything under the sun in stability characterization, including thermal unfolding, particle sizing, and aggregation.


♦ Prometheus NT.Plex

Your best bet when you want an easy path to automation. Get thermal unfolding and chemical denaturation with optional backreflection for aggregation evaluation.


♦ Prometheus NT.48

Start here for thermal unfolding and chemical denaturation with optional backreflection for aggregation evaluation.



Automated operation for unattended stability characterization



Prometheus NT.Plex plus NT.Robotic Autosampler

Add NT.Robotic Autosampler to transform Prometheus NT.Plex into a system that increases your throughput with unattended operation. It’s a stand-alone, all-inclusive system with robotic arm, computer, and monitor. Analyze up to 1,536 samples for hands-free characterization of thermal unfolding and chemical denaturation in a single experiment. Incorporate plate temperature control to keep your samples at 4 to 20 °C while they wait in the queue. And, include aggregation detection optics to get you complete stability characterization.












Tackle challenging stability characterizations with multiple technologies in one instrument

Prometheus utilizes a number of technologies to characterize thermal unfolding, particle sizing, and aggregation. Each one tells a different story about stability, and is used for a variety of applications. But they have one thing in common — they are all label-free.



When is it useful?


What does it do?

For monitoring protein purification, evaluating target-ligand complexes before affinity-based screening, or monitoring stability during biologics formulation, developability, production, and manufacturing


It measures thermal unfolding or chemical denaturation under native conditions and label-free by detecting changes in its intrinsic fluorescence during a thermal ramp or in the presence of a chemical denaturant



When is it useful?


What does it do?

For screening recombinant proteins expressed from different constructs, biologics formulation screening, sample optimization for biophysical assays, and characterization of self-interactions


It detects molecule aggregates to determine the size of proteins over a wide concentration range




When is it useful?


What does it do?

Anytime you’re looking at aggregates larger than 12.5 nm radius


It determines aggregation status by measuring light intensity loss due to scattering



When is it useful?


What does it do?

For determining if complexes have formed, measuring how much genetic material a viral vector contains, or extrapolating how proteins will interact with themselves at high concentrations.


It looks at the overall average scattering intensity of your solution and correlates that to the molecular weight of your particles.


Parameters that characterize protein stability and particle sizing

Do you wonder what attributes are used the most when looking at protein stability? These are the parameters researchers look at when characterizing the propensity of proteins to unfold and aggregate in response to binding events, self-interactions, temperature changes, or the presence of certain chemicals


Thermal Stability



The unfolding transition temperature is the point at which 50% of the protein is unfolded. Proteins with a higher Tm are more stable because a greater input of energy is required to reach the unfolding transition. Because Tm is an accurate and established metric for assessing protein stability, it’s an essential parameter to determine.




The detectable temperature at which a protein begins to unfold. Particularly in temperature-sensitive situations, it’s important to understand when a protein will denature and lose its activity.



See how you can characterize both thermal unfolding and chemical denaturation with nanoDSF

Prometheus does it best when it comes to characterizing protein unfolding. In both thermal and chemical denaturation studies, even the most subtle unfolding events are easily seen that other traditional systems simply can’t detect. Best of all, results are not compromised by aggregates in solution. Get higher quality results and gain the ability to make better decisions.


Prometheus monitors the intrinsic fluorescence signal of proteins as a measure of their folding state. Fluorescence intensity at a single wavelength (350 or 330 nm) or the F350/330 ratio are plotted against increasing temperature or concentrations of a chemical denaturant to determine the Tm or Cm of a protein.




     Prometheus offers precise, high-resolution results in protein denaturation studies


Chemical denaturation

 Cm or C50 


The concentration of a denaturant that causes 50% of proteins to unfold addresses the relationship between chemical denaturants and proteins. This parameter is useful when studying chemical effects, or when protein stability is tested using denaturants.


ΔG and ΔΔG


The Gibbs free energy of protein unfolding, is a thermodynamic measure of the likelihood a folding event may occur. For a given event, proteins with more negative ΔG are more likely to fold. ΔG also relates protein folding to changes in enthalpy and temperature. Changes in the Gibbs free energy are measured with ΔΔG, which shows the relative stability of a protein at various concentrations.


Particle sizing




The hydrodynamic radius tells how large or small a protein is in a solvated state, making it a biologically relevant parameter since it considers the protein size in the context of its environment. Monitoring the rH is a simple way to identify the oligomeric state of a protein in a preparation and can be implemented during production to identify relevant fractions of purified protein or to check batch to batch consistency of formulations.




The polydispersity index represents the distribution of size populations within a given sample. The numerical value of PDI ranges from 0.0 (for a perfectly uniform sample with respect to the particle size) to 1.0 (for a highly polydisperse sample with multiple particle size populations)


kD and D0


The diffusion interaction parameter (kD) identifies the onset of protein unfolding and its impact on colloidal stability. Positive kD values are a sign of repulsive intermolecular interactions, while negative values indicate attraction. It can be utilized to compare different protein formulations, and use it to select more stable biomolecules.
The diffusion constant at concentration = 0 (D0) is determined from the kD analysis and represents the theoretical diffusion constant at concentration = 0.


See how you can select the best buffer formulation using DLS


Compare size distribution, mean rH, and PDI to identify optimal buffer conditions for your sample and assess sample homogeneity before performing other time-consuming biophysical assays













This is the temperature at which proteins exhibit a tendency to aggregate. Typically, the onset of turbidity correlates with the unfolding of proteins and can be a useful parameter for comparing colloidal stability.


See how a team at Boehringer Ingelheim used thermal unfolding and aggregation to predict the stability of their candidate molecules

Being confident in what to do next is key to doing great research. The formulation development group at Boehringer Ingelheim realized that having accurate, precise, and high-quality data for both thermal unfolding and aggregation is key to better predicting stability, developability, and longer-term storage of their antibody candidates.


Prometheus provides comprehensive results for any type of proteins — small or large molecules, biologics, enzymes, antibodies, ADCs, and membrane proteins — and is especially good at screening buffer influences or testing formulation and storage conditions. Make better decisions based on more complete stability results.



Prometheus provides comprehensive stability results


Conformational stability (or thermal unfolding) doesn’t always correlate with colloidal stability (or aggregation). Here, the lowest pH destabilizes the mAb, yet reduces its propensity to aggregate. Conversely, the highest pH stabilizes the mAb but causes an increase in the level of aggregation. Therefore, selecting a pH between the two extremes may be the better formulation compromise to use.


Particle weight determination

Molecular weight

Your protein’s molecular weight will change if it binds to another protein or molecule. You can also determine if a gene therapy vector contains genetic material or not.


Second virial coefficient, B22

If the B22 of your protein is positive, that indicates it prefers to be solvated – that is, it is less likely to self-interact, and therefore aggregate. Measure your sample at low concentrations to predict how it will behave after scale-up.



What researchers look for when selecting an instrument to characterize stability

Characterizing a candidate molecule’s stability with an instrument that has high specificity, high sensitivity, a broad limit of detection, is very accurate as well as very precise, and produces high resolution data makes a huge difference. It allows you to identify or anticipate structural liabilities that render your molecule non-manufacturable. It also helps to minimize costly mistakes like progressing a candidate that will ultimately not make it to the clinic.

With Prometheus, never doubt the quality of your measurements and move forward with confidence knowing your decisions are derived from the highest quality data.


High level of specificity


Distinguish the signal of your biologic from the signal of the buffer or matrix

High level of sensitivity


Identify samples and domains with subtle signals

Broad limit of detection


Measure samples at both clinically and developmentally relevant concentrations, including both high and low concentrations



Report true Tms



Provide reliable results that consistently return the same value with little error consistent and reliable results with small standard deviations or errors that do not vary from day-to-day

High resolution


Detect multiple unfolding events discriminates domains with similar Tms



Software that gives you clear and actionable results

Wondering how long it will take your team to learn to operate a new instrument and to use its software can trigger stress and a feeling of uncertainty. Not if your instrument is a Prometheus. No matter which model you choose, learning how to use it will be effortless with step-by-step guidance through each experiment, the automatic analysis of your data, and much more.


   Panta Control

Because Prometheus Panta measures more parameters simultaneously, Panta Control software is built with more functionality and intuitive user experience. Get automatic determination of the parameters you care about the most. When you decide to measure multiple parameters, queue them in your preferred order for flexibility that adapts to your schedule. Learn more



Precisely determine at which temperature 50% of your protein unfolds (Tm) and when aggregation begins (Tturbidity) with PR.ThermControl. Since assay setup is only 3 steps and there’s few experimental parameters to set, you’ll be off and generating data quicker than you imagine. Results are automatically generated and color coded so you can easily visualize the results. Learn more



Use PR.ChemControl when you want to evaluate the chemical stability of a protein. It only takes 40 seconds to get data on the energetics of folding (ΔG) and the denaturant concentration at which 50% of the protein is unfolded (Cm). It’s great for getting quick answers on chemical treatments or conditions that influence protein unfolding and stability.



Are you searching for more details on the rates of unfolding and refolding? Maybe you need to run more advanced thermal profiles such as isothermal measurements or thermal cycling. Your ability to customize thermal treatment assay conditions is one of the best reasons to use PR.TimeControl. Feel free to choose between isothermal measurements, incremental temperature cycling or temperature stepping for those advanced protein stability measurements you’ve always wanted to do.


PR.Stability Analysis

If making more precise conclusions about the stability of your proteins is your ultimate goal, then there’s no better way to do that than with PR.Stability Analysis software. Use it to quickly visualize and identify any key trends by sorting, filtering or plotting data. Choose to merge replicates or compare runs, then conveniently export your results in ready-made formats for bookkeeping, presentation or publication. Learn more


21 CFR part 11

Prometheus PR.ThermControl and PR.AutoThermControl software support 21 CFR Part 11 compliance for regulated environments. They provide electronic signatures, audit trail with record of all activities, user permissions, and access management.




Get consistency with high performance consumables


Prometheus-CapillariesYou don’t have to worry about a long list of consumables to run your stability assays. All you’ll need are capillaries — but not just any, because only high quality consumables will deliver consistent results. Prometheus capillaries are manufactured using the same stringent protocols used for diagnostic-grade ones to ensure the best results. They come individually or in chips.











Want to learn more about Prometheus? View Brochure


For more information or ordering the products, please email us at cs@medikonia.com.

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