Analytical Ultracentrifugation (AUC) Technology

What is Analytical Ultracentrifugation (AUC)?

AUC is a powerful tool for studying the size and shape of protein macromolecules in solution. Compared to other methods for analyzing macromolecules, AUC is able to characterize samples in their native state under biologically relevant solution conditions, providing pertinent information about the molecular weight, and hydrodynamic and thermodynamic properties of proteins or other macromolecules. In addition, the technique can be used to study virtually any type of molecule or particle, with a wide range of concentrations.

Analytical Ultracentrifugation (AUC) Principle

AUC operates on the principle that macromolecules in solution will settle at a certain speed when subjected to centrifugal force, including two main modes of operation: sedimentation velocity (SV-AUC) and sedimentation equilibrium (SE-AUC).

In SV-AUC and SE-AUC analysis, the time-dependent concentration curve of the macromolecule is measured using one or more detection methods such as UV absorbance and fluorescence detection. Subsequently, computer algorithms process these time-dependent concentration profiles to fit the appropriate mathematical model. During the analysis, the speed of movement of the boundary provides information about the size, shape, and mass of the macromolecule. The shape of the boundary layer provides quantitative information about the distribution of macromolecules of different sizes in the solution, such as the number of dimers/aggregates.

The difference between the two is that the SE-AUC technique is sensitive to the mass of the macromolecule, but not to the shape, and can be used to determine the absolute molecular weight of aggregates present in the solution. It should be noted that the experiment was performed at a lower speed, where back diffusion from the bottom of the cell (high concentration) to the top (low concentration) becomes significant.

Fig. 1 Basic analytical ultracentrifugation experiments. (Cole JL, et al., 2008)

Our Services Related to Analytical Ultracentrifugation (AUC)

With decades of experience supporting protein/peptide biopharmaceutical development and manufacturing, our team of highly qualified experts can provide a range of AUC-related services to accelerate the implementation and success of your project.

Our laboratories are equipped with the most advanced AUC technology with first-class detection methods and professional technicians to provide the most professional AUC analysis services for all stages of protein/peptide drug development and production - from early research to late-release testing under GMP.

We use AUC technology to focus on explaining aggregation problems in protein/peptide production and manufacturing processes. By exploring the mathematical correlation between aggregate sedimentation behavior and fluid dynamic properties, our scientists can determine many key parameters of protein/peptide drug substances or drug products, including aggregation propensity, stability, aggregate size distribution, conformational changes, etc. These parameters provide a range of critical information that can be used to optimize production processes, improve product quality, and ensure the safety and efficacy of protein/peptide drug products.

Our AUC-related services include:

• Protein/peptide biopharmaceutical characterization (molecular weight, size distribution, diffusion coefficient, sedimentation coefficient, etc.).
• Detection of aggregates in protein samples and quantification of aggregate concentration.
• Protein-protein interaction studies.
• Protein/peptide nanoparticle delivery systems characterization.
• Elucidation of protein higher-order structure to support similarity studies
• Establishment of the native state of proteins, monomers, dimers, etc.

Our Equipment

At CD Formulation, our analytical ultracentrifuges are equipped with a variety of detectors, including a fluorescence detector, UV-Vis detector, and Rayleigh interferometer optics.

• The fluorescence detector detects compounds that fluoresce after excitation, such as fluorescently labeled proteins or nucleic acids. This detector provides greater sensitivity and selectivity and is suitable for detecting low concentrations of target substances.
• UV-Vis detector can detect light-absorbing protein substances in solution. This is because they usually absorb light in the UV at 280 nm.
• Rayleigh interferometer optics monitor changes in refractive index and can be used to track protein/peptide compounds that do not absorb in the UV or visible range.

Advantages of Our Analytical Ultracentrifugation (AUC) Technology

• High resolution: Provides high-resolution data on the size, shape, and molecular weight of protein/ peptide and nanoparticles.
• Versatility: AUC can be used to analyze a wide range of samples, including proteins, nucleic acids, viruses, nanoparticles, and polymers.
• Label-free analysis: AUC does not require any labeling or modification of samples, allowing for the analysis of native, unaltered protein.
• Minimal sample preparation: AUC typically requires minimal sample preparation, making it a quick and efficient technique for analyzing biomolecular interactions.
• Quantitative analysis: AUC can provide quantitative data on the stoichiometry, binding constants, and thermodynamic parameters of biomolecular interactions.
• Accurate measurement of sedimentation coefficients: AUC can accurately measure sedimentation coefficients, providing information on the size and shape of macromolecules.
• Real-time monitoring: AUC can provide real-time monitoring of molecular interactions, allowing for the study of complex biological processes.
• High sensitivity: AUC can detect low concentrations of biomolecules, making it a valuable tool for studying protein-protein interactions and protein-ligand interactions.
• Wide range of experimental conditions: AUC can be performed under various experimental conditions, including different buffers, temperatures, and pH values, allowing for the study of biomolecules in physiological conditions.
• Compatibility with other techniques: AUC can be used in conjunction with other analytical techniques, such as mass spectrometry and fluorescence spectroscopy, to provide comprehensive information on biomolecular interactions.

Custom Analytical Ultracentrifugation (AUC) Services

Why Choose Our Analytical Ultracentrifugation (AUC) Technology?

• We have a team of experts with rich experience in performing AUC analytical method development and validation.
• We have accumulated decades of expertise and successful project experience using AUC technology to support protein/peptide biopharmaceutical development.
• With our efficient AUC analysis tool, we can perform detailed characterization of different aspects of different types of protein/peptide therapeutics.
• We provide in-depth information on molecular weight, aggregate content, impurities, and stability of protein/peptide drugs.
• We provide flexible experimental design and customized solutions.
• Fast turnaround time: detailed technical reports are provided within 5-7 days.

Publication

CD Formulation aims to provide a powerful analytical tool for studying the size and shape of protein macromolecules in solution. Please feel free to contact us to speak with our team of scientists and learn how our AUC technology can support your product development.

How It Works

STEP 1

Submit a service request describing your specific research or development need.

STEP 2

We'll email you to provide your quote and confirm order details if applicable.

STEP 3

Execute the project with real-time communication, and deliver the final report promptly.