Nano Differential Scanning Fluorimetry

Our Services
Nano Differential Scanning Fluorimetry

Method Introduction

Nano differential scanning fluorimetry (nanoDSF / nDSF) is a fast and low-volume technique to determine the melting temperatures of proteins using their intrinsic fluorescence.

In nanoDSF, a protein in solution is exposed to a temperature gradient that will lead to the unfolding of the protein. The intrinsic fluorescence of the protein, mainly originating from the aromatic side chains of tyrosine and tryptophan residues, is examined. Upon unfolding, the environment of those residues will alter because they become exposed to the solvent, and thus, their fluorescence spectrum will change. The relation between the fluorescence changes and the temperature gradient can be used to obtain a so-called “apparent” melting temperature (Tm). The melting temperature is called “apparent” because the thermal denaturation process can very often lead to aggregation at high temperatures and affect the measured Tm value. Moreover, the Tm value obtained by nanoDSF may differ from the one obtained by differential scanning calorimetry (µDSC) due to the different parameters measured (global unfolding for µDSC and changes in the environment of the aromatic amino acids due to the tertiary folding in nanoDSF). Nonetheless, nanoDSF can also provide information about the nature of the unfolding process, indicating if it involves a cooperative (two-state) or complex unfolding transitions, like observed in multi-domain proteins.

Need more information? Follow the links below and contact our experts with your questions today.

Applications

nanoDSF allows rapid analysis of multiple samples in parallel under identical conditions, e.g., within a formulation development study using different stabilizing excipients, thus being a valuable tool for formulation screening studies.

Besides the small sample volumes required (as low as 10 µL), nanoDSF offers much shorter analysis times and is easier to set up than DSC. Thus, it is a highly valuable alternative to µDSC, particularly in early research and development.

Quality and Biosafety Level

We provide all our analytical services with the highest quality standards. Experienced scientists carry out each project, and a scientific reviewer comprehensively checks every report or data presentation.

We offer this technology with the following quality and biosafety levels:

R&D level

We offer this method under R&D. Our GRP system assures the highest-quality research standards.

Up to biosafety level 1

This method can be applied to proteins, nucleic acids, and most viral vectors, including AAVs and more.

Nano Differential Scanning Fluorimetry Frequently Asked Questions (FAQs)

  • NanoDSF is a biophysical method used to measure the thermal stability of proteins by monitoring their intrinsic fluorescence during controlled heating. It determines the apparent melting temperature (Tm) of a protein as it unfolds.

  • nanoDSF detects fluorescence changes rather than heat flow. While both methods assess protein stability, nanoDSF requires less sample volume, offers faster analysis, and is easier to set up compared to µDSC making it highly efficient for early-stage formulation screening.

  • nanoDSF is suitable for proteins, peptides and some viral vectors, such as adeno-associated viruses (AAVs). It requires very small sample volumes—as low as 10 µL—making it ideal for limited or high-value materials.

  • nanoDSF helps identify stabilizing excipients, compare formulations, and screen for optimal buffer conditions. It can also reveal whether unfolding is a simple two-state transition or a complex multi-domain process.

  • One of the reasons why the melting temperature (Tm) obtained via nanoDSF is considered “apparent” is because it’s based on changes in intrinsic fluorescence. This may not directly align with Tm values from µDSC, as fluorescence and heat transitions don’t always correlate perfectly.

  • Yes. nanoDSF supports parallel analysis of multiple samples under identical conditions, making it highly effective for high-throughput stability screening during early development phases.

  • Coriolis offers nanoDSF under research-grade (R&D) conditions with Good Research Practice (GRP) standards and supports biosafety level 1 (BSL-1) applications. Every report is reviewed by experienced scientists to ensure data integrity.

  • nanoDSF complements other techniques such as circular dichroism (CD), µDSC, dynamic light scattering (DLS), and FTIR spectroscopy. It’s often used early in formulation development or as part of higher-order structure analysis.

  • nanoDSF is fast, cost-efficient, and ideal for low-volume, high-value protein samples. It delivers valuable thermodynamic information with minimal setup, enabling rapid decision-making in formulation and stability studies.

  • Yes. Under BSL-1 conditions, nanoDSF can be applied to certain viral vectors, including AAVs, to evaluate thermal stability and assess formulation robustness.

Analytical Method Development, Qualification and Validation

For common sample types, we can often apply standardized methods with little setup effort. However, when needed, our experienced analytical experts create or optimize custom methods tailored to your active pharmaceutical ingredient, product type and development phase.

Method Development

Our method development approach tailors sample preparation, method settings and data analysis to the needs of your project and sample.

We include representative samples and, where available, suitable reference standards and stressed/degraded materials, allowing our analytical scientists to design a highly suitable, stability-indicating, robust and repeatable method. Upon request, we will compile a detailed description of the method for your records.

Method Qualification

Method qualification is the initial assessment of an analytical procedure’s performance to show its suitability for its intended purpose.

During method qualification, our analytical scientists perform documented testing demonstrating that the analytical procedure meets criteria in several categories. Criteria may include factors such as repeatability, specificity and robustness. We compile a qualification plan and report, including all relevant data.

Method Validation

Under GMP conditions, method validation confirms that an analytical procedure’s performance suits its intended purpose. Depending on the method’s scope, a broad range of method characteristics, such as specificity, accuracy, precision, limit of detection/limit of quantification (LOD/LOQ), linearity and range, is considered.

During method validation, our analytical scientists perform documented testing demonstrating that the analytical procedure consistently produces a result that meets the predetermined acceptance criteria. We compile a validation plan and report that includes all relevant data.

Depending on the development phase, a fit-for-purpose validation approach can be offered, adjusting the validation required efforts in a phase-appropriate way to meet the method’s needs.

Method Verification

Compendial method verification confirms that a compendial method (e.g., from Ph. Eur. or USP) is suitable and reliable for its intended purpose under the specific conditions of the laboratory.

Unlike full method validation, compendial method verification is often considered a partial validation since the method has already undergone extensive testing and validation during its inclusion in the compendium. The extent of method verification depends on the type of method.

During method verification, our analytical scientists perform documented testing demonstrating that the developed analytical method performs adequately for the specific product or matrix being tested and within the specific laboratory where the method will be employed.

Talk to Our Experts or Request a Quote

Our expert team is ready to answer your questions and guide you to the services best suited to your program’s modality, stage and challenge. If your needs are well-defined, we’ll begin the quotation process.

Description

Related Services

Higher Order Structure – Overview Circular dichroism (CD) Differential scanning calorimetry (µDSC) Dynamic Light Scattering (DLS) Fluorescence spectroscopy Fourier transform infrared (FTIR) spectroscopy High-performance size-exclusion chromatography (HP-SEC) Liquid chromatography coupled mass spectrometry (LC-MS)