How Analytical Ultracentrifugation (AUC) Works

Introduction

Analytical ultracentrifugation (AUC) is a precise and versatile technique for the biophysical characterization of macromolecules in solution. By applying high centrifugal forces to the sample, the Optima AUC monitors the hydrodynamic and thermodynamic behaviors of proteins, nucleic acids, viruses and other particles in their native state. Providing critical insights into molecular weight, size distribution, shape, and interactions, it is an indispensable tool in fields such as biochemistry, molecular biology, and nanotechnology.

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Key Benefits of AUC

Stand-alone technique performed in a matrix-free environment
It is dye-free and label-free
Broad buffer compatibility / limited buffer constraints
Valuable for studying molecular interactions and complex systems in solution
First principle characterization technique; no standards required

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A scientist works with a Beckman Coulter Optima AUC analytical ultracentrifugation instrument in a laboratory.

Applications of Analytical Ultracentrifugation

Whether you are studying proteins, nucleic acids, polymers, virus-like particles, or nanoparticles, Optima AUC offers unparalleled flexibility and precision. It can detect and characterize macromolecular oligomeric states, heterogeneity and conformational changes in response to biological or environmental variations such as pH and temperature. AUC provides unparalleled flexibility with the ability to measure particles from small peptides to large viruses over a broad concentration range and a wide range of solvents.

The Optima AUC is crucial for determining the stoichiometries, molecular masses, and sizes of macromolecules, as well as for analyzing their reversible and non-reversible interactions. Additionally, AUC is invaluable for distinguishing particle loading states, making it an indispensable tool in molecular biology and biochemistry.

Discover how AUC can enhance the characterization of your macromolecules and elevate your research.

Viral Vectors

AUC in mAbs

AUC in LNP

AUC in Extracellular Vesicles

AUC in Protein Research

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Optima AUC: Multiple Methods in One

Analytical ultracentrifugation is a broadly applicable and information‐rich technique for investigating macromolecular characteristics, such as size, shape, stoichiometry, and binding properties, in their native state under biologically relevant conditions - capabilities often lacking in orthogonal technologies.

The Optima AUC is equipped with two integrated optical detection systems: the UV-Vis absorbance optical system, which allows for the detection of absorbing analytes in solution between 190-800 nm, and the Rayleigh Interferometer, which can be used to monitor and detect any analytes that differ between the sample and reference solutions.

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A scientist in a lab coat and gloves stands next to the Optima AUC instrument.

Sedimentation Velocity, Sedimentation Equilibrium, and Density Gradient Equilibrium

AUC is extremely versatile, as it can employ several orthogonal methods. These methods allow the researchers to tailor their experiments to meet specific needs and achieve precise results across different applications and concentration ranges.

Some key methods include Sedimentation Velocity (SV), Sedimentation Equilibrium (SE), and Density Gradient Equilibrium (DGE). Each method offers distinct insights: Sedimentation Velocity provides high-resolution hydrodynamic information about molecular size and shape; Sedimentation Equilibrium offers thermodynamic details like accurate molar masses; and Density Gradient Equilibrium separates particles by buoyant density within a gradient, which can reduce sample requirements.

Additionally, a recent innovation possible with the Optima AUC, multiwavelength AUC, can extract spectral properties of the analytes in SV and DGE experiments significantly enhancing interaction studies. To learn more about these methods and how they can enhance your research, follow the links below.

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