Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC) Data Evaluation Approaches

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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. 

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.

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.


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SV-AUC Data Evaluation Approaches

Nowadays, sedimentation velocity experiments (SV-AUC) are evaluated by finite element methods. As Beckmann Coulter, the worldwide leading builder of AUC instrumentation, never developed a data evaluation software for analytical ultracentrifugation, the academic community took over the task. Several different approaches were taken to evaluate the data. Therefore, a large number of different programs for SV-AUC data evaluation are available. ZentriForce Pharma offers two main data evaluation services (Ultrascan and Sedfit) for routine measurements. Even though we focus on these two programs our scientists are experienced with a wide range of other software as well. Should you have a different data evaluation approach in mind regarding your product, we are more than happy to assist you.   

Sedfit c(s)

Sedfit c(s) data evaluation approach is the most commonly used data evaluation in the pharmaceutical industry. It is a fast and easy to use data evaluation approach. The algorithm can be applied to calculate sedimentation coefficient distributions with information such as molecular weight and partial concentrations of the appearing species. The software, however, has some drawbacks. The program shows user dependent effects on the results. At ZentriForce we reduced this dependence to a minimum. However, it is impossible to get completely user-independent results. Additionally, the automatic peak detection of the program is not 100% reliable. In some cases, different numbers of peaks are detected. The c(s) is also limited to a global frictional ratio (shape information). Sedfit c(s) is a good algorithm for fast and cost sensitive data evaluation. For most applications Sedfit c(s) is an acceptable choice.  However, for larger measurement campaigns (e.g. biosimilarity studies or stability studies) we recommend the use of the Ultrascan PCSA-MC algorithm. 

Ultrascan PCSA-MC

Ultrascan data evaluation requires the use of a computer cluster to run data evaluation. In academia, public computer clusters (e.g. Jülich) are used. However, for a CRO such as ZentriForce Pharma, that is not an option, as data confidentiality has highest priority. Therefore, we decided to set up our own computer cluster. This way we can offer to evaluate SV-AUC data using the Ultrascan PCSA-MC algorithm. The algorithm enables us to test for different correlations between sedimentation coefficient and frictional ratio (e.g. increasing sigmoidal, linear) and not only one global frictional ratio. Additionally, we are able to refine the results by additional Monte Carlo Simulation. For the peak definition we can define distinct integration ranges. That is how we can ensure to always use identical data evaluation (not possible in Sedfit c(s)) The combination of these factors makes the PCSA algorithm more robust and reliable. Therefore, we recommend to use it for biosimilarity studies.    

We are currently working in a cooperation with the Ultrascan team to develop a GMP-ready AUC data evaluation approach. As we see the rising need for SV-AUC services on a GMP level we are working on the possibility to offer GMP-qualified SV-AUC as soon as possible.   

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. 

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.

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.

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