Analytical Ultracantrifugation Theory

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.

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. Follow this link to get more information about SE-AUC theory.

Even though SV-AUC is with its more than 100 years of history a by far older technique than most others currently used within biopharmaceutical development. It is one of the most complex in regards of data evaluation. We have setup our own supercomputer cluster to be able to offer you several different data evaluation approaches, without the need of your data leaving our premises. We currently offer Sedfit and Ultrascan data evaluations. However, if we see the need to use a different software for your very molecule we have the knowledge base to do so. Please follow the link for more information. 

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.


General topics:

ZentriForce Pharma was founded as a service provider that specializes in hydrodynamic techniques for biopharmaceutical development. The focus lies on gene therapy products and biosimilarity studies. Follow the link to our company page for further information.

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ZentriForce Pharma implemented a strict quality system to assure the highest tractability of results generated at our facility. We have a wide range of quality related standard operation procedures (SOPS) in place to assure the consistency and quality of our laboratory operations. Please follow the link to a description of our quality system. 

ZentriForce Pharma is your expert in analytical services, with focus on hydrodynamic techniques. Please follow the link to find out about our analytical services. 

ZentriForce Pharma offers a wide range of analytical techniques. Please follow the link to our analytical portfolio. 

ZentriForce Pharma is not merely an expert in analytical development. Our scientist are actively involved in further advances in the theory of hydrodynamic techniques. Follow the link to a detailed description of the theory behind our analytical services, including some of our own research

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Analytical Ultracentrifugation (AUC)

With roughly 100 years of history, analytical ultracentrifugation (AUC) is a comparably seasond yet still well-established method.  Although AUC had initially been developed for the analysis of nanoparticle size distributions, the focus quickly shifted toward biological applications such as biopolymers.

Due to time consuming photographic data acquisition in the early instruments and the upcoming of new methods for biopolymer analysis, such as HPLC, AUC lost its importance as one of the leading analytical techniques for biopolymers during the middle of the last century.

Since the introduction of a new generation of analytical ultracentrifuges, namely the Optima XL-A by Beckman Instruments (Palo Alto, USA) in the 1990’s, a renaissance of analytical ultracentrifugation has taken place. However, nearly 30 years without significant technical improvements took their toll on the impact of AUC in biopharmaceutical development.

In 2017 the new state-of-the-art instrument, Optima AUC, was released. The capacities of the Optima might lead to a rebirth of AUC.  

Sedimentation Velocity Analytical Ultracentrifugation

The sedimentation velocity measurement is nowadays the standard AUC experiment. It is carried out at high centrifugal velocity and provides the easiest way to obtain sample composition and species characteristics (e.g. molecular weight and frictional ratio). In a sedimentation velocity experiment the solutes are separated according to three parameters, which are shape, size and density. For more detailed information, please visit our SV-AUC theory page

Sedimentation Equilibrium Analytical Ultracentrifugation

While sedimentation velocity is the standard AUC experiment, sedimentation equilibrium analytical ultracentrifugation (SE-AUC) equally offers valuable information on an analyte.  SE-AUC is a first principle method for the determination of the molecular weight and therefore an excellent choice to analyze your API in solution. For more detailed information,  please visit our SE-AUC theory page.  

Comparison Optima AUC vs ProteomLab XL-A

Nearly 30 years have passed since the Proteomlab Xl-A was introduced by Beckmann Coulter. The technical advances from the early 90’s to today bear a potential for much higher data quality. The technical differences between Optima AUC and ProteomLab XL-A can be found in the table below.

The Optima AUC provides a three times higher data density and measures much faster (roughly four times faster). Therefore, much more data can be collected within a comparable measurement time. This is especially useful for the quantification of larger aggregates (normally only a small number of scans are available including information of aggregates).

The higher wavelength accuracy allows the parallel collection of serval UV signals (e.g. 260 nm and 280 nm for adeno-associated virus (AAV) products). The ration of two wavelength can be a valuable information for the characterization of species observed. The changed specifications of the CCD camera allow the measurement of higher concentrated samples with interference optics.

Table 1: Specifications Optima AUC vs ProteomLab XL-A

PROTEOMELAB XL-A / XL-I

Instrument

OPTIMA AUC

ABS: 90sec/cell INT: 5 sec/scan

Fastest data acquisition rate

 ABS: <20 sec/cell INT: <5sec/scan

 +/- 3 nm

Wavelength precision

 +/- 0.5 nm

30 μm

Lowest radial resolution

10 μm

2048 x 96 pixels

CCD camera specifications

 2048 x 1088 pixels

 

 

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.

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. Follow this link to get more information about SE-AUC theory.

Even though SV-AUC is with its more than 100 years of history a by far older technique than most others currently used within biopharmaceutical development. It is one of the most complex in regards of data evaluation. We have setup our own supercomputer cluster to be able to offer you several different data evaluation approaches, without the need of your data leaving our premises. We currently offer Sedfit and Ultrascan data evaluations. However, if we see the need to use a different software for your very molecule we have the knowledge base to do so. Please follow the link for more information. 

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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