Sedimentation Equilibrium Analytical Ultracentrifugation (SE-AUC)

Related topics:

Sedimentation velocity analytical ultracentrifugation (SV-AUC) is the gold standard in aggregation analytics. The importance of SV-AUC results is growing, especially in biosimilarity studies, and also within the development of gene therapy products.   ZentriForce Pharma offers SV-AUC services for a wide sample spectrum. For more information please follow the link.

Sedimentation velocity analytical ultracentrifugation (SV-AUC) method is the established standard experiment when it comes to AUC. It is carried out at high rotational speed and the easiest way to obtain sample composition and species characteristics (e.g. molecular weight and frictional ratio). Follow this link to get more information about SV-AUC theory.

Light scattering is a well defined effect. The physical properties of light scattering are used for several applications, such as dynamic and static light scattering. Please follow the link to a detailed description of light scattering theory. 

In the last couple of years, the interest in the determination of predictive parameters within formulation development has grown. Interactions parameters provide valuable information for the prediction of molecular stability. These values are used in a “design of experiment” (DOE) approach to assist formulation developments. Please follow the link for more information.

With the increasing interest in gene therapy products also the interested in AF4 is growing. However, AF4 methods are comparably complex to develop. While most people use AF4 only as separation technique. We are experts in AF4 theory and use the theory to get more information out of the technique (e.g. particle size distributions). If you want to get more information about AF4 theory please follow this link.


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Sedimentation Equilibrium Analytical Ultracentrifugation (SE-AUC)

While sedimentation velocity is the standard AUC experiment, sedimentation equilibrium analytical ultracentrifugation (SE-AUC) nonetheless offers valuable information on an analyte.  SE-AUC is a first principle method for the determination of the molecular weight. Therefore, it is the method of choice to determine the molecular weight of your API in solution. The measurement, however, takes much longer than SV-AUC measurements. While SV-AUC measurements are finished after a maximum of 24 h, SE-AUC measurements can easily take a full week to reach an equilibrium.

SE-AUC Theory

In an SV-AUC experiment the rotation speed is high enough to reduce the back diffusion of the analyte to a low level. In the case of SE-AUC the rotation speed is reduced to a level that allows significant back diffusion.  SE-AUC experiments are run at low speed until an equilibrium between sedimentation and back diffusion is reached. As we reach an equilibrium, the shape of the molecule no longer influences its behavior and its molecular weight can be assessed from the position and slope of the resulting concentration gradient. In most cases, it is recommended to measure several different rotational speeds and at different concentrations. Two approaches for the determination of sedimentation equilibrium are available. In a high-speed sedimentation equilibrium, the rotational speed is chosen such that, even though an equilbrium is reached, the concentration gradients start below the meniscus. The second option a low speed SE-AUC experiment, in which a lower rotational speed is applied. The resulting concentration gradient is much shallower. Which of these two experiments should be applied depends on the sample properties.  

Additional Applications for SE-AUC

SE-AUC has a particularly interesting advantage over SV-AUC. In the case of SE-AUC, concentration dependence is not as problematic as for SV-AUC samples. In most cases it can be applied to samples at their original concentration. However, in this case a dilution series is recommended to determine the concentration dependence of the molecular weight. The concentration dependence of the molecular weight is influenced by the second osmotic virial coefficient (B22), which quantifies repulsive molecule – molecule interactions. For more information please visit our interaction parameter determination page

Sedimentation velocity analytical ultracentrifugation (SV-AUC) is the gold standard in aggregation analytics. The importance of SV-AUC results is growing, especially in biosimilarity studies, and also within the development of gene therapy products.   ZentriForce Pharma offers SV-AUC services for a wide sample spectrum. For more information please follow the link.

Sedimentation velocity analytical ultracentrifugation (SV-AUC) method is the established standard experiment when it comes to AUC. It is carried out at high rotational speed and the easiest way to obtain sample composition and species characteristics (e.g. molecular weight and frictional ratio). Follow this link to get more information about SV-AUC theory.

Light scattering is a well defined effect. The physical properties of light scattering are used for several applications, such as dynamic and static light scattering. Please follow the link to a detailed description of light scattering theory. 

In the last couple of years, the interest in the determination of predictive parameters within formulation development has grown. Interactions parameters provide valuable information for the prediction of molecular stability. These values are used in a “design of experiment” (DOE) approach to assist formulation developments. Please follow the link for more information.

With the increasing interest in gene therapy products also the interested in AF4 is growing. However, AF4 methods are comparably complex to develop. While most people use AF4 only as separation technique. We are experts in AF4 theory and use the theory to get more information out of the technique (e.g. particle size distributions). If you want to get more information about AF4 theory please follow this link.

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