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Analytical Ultracentrifugation

A precise and powerful tool for studying the size and shape of macromolecules in solution

Analytical Ultracentrifugation (AUC) Services

Part of KBI Biopharma's Rapid Analytics Biophysical Characterization Core


Analytical ultracentrifugation (AUC) is a precise and powerful tool for studying the size and shape of macromolecules in solution. 

AUC is particularly important for characterizing protein aggregation and determining the native state stoichiometry (quaternary structure). AUC is also commonly used to show comparability of higher order structure (HOS). 

Using the newer Beckman Optima AUC, the KBI Biopharma Rapid Analytics Biophysical Core has established a multi-wavelength approach that identifies and quantifies various fill states in viral systems like adeno-associated virus (AAV) and Adenovirus (Ad) systems for applications like gene therapy. 

We have also introduced a new sedimentation equilibrium protocol that allows us to measure the weight-average mass of highly-concentrated protein formulations (up to ~200 mg/mL) requiring only ~20 µL of sample per measurement. 

KBI Biopharma's Rapid Analytics Biophysical Core has completed hundreds of AUC projects on proteins, peptides, nucleic acids, viruses, and other macromolecules. All of the analytical ultracentrifugation experiments, data interpretation, and data review are done by recognized AUC experts with extensive experience in AUC applications and data analysis with many publications involving AUC. 

We use six analytical centrifuges at the Rapid Analytics Biophysical Core: two Beckman-Coulter XL-I instruments equipped with both absorbance and Rayleigh interference (refractive index) detection, a Beckman-Coulter XL-A for absorbance data only, and three Beckman Optima AUCs for interference and absorbance optics. The Optima is a state-of-the-art AUC instrument with improved signal to noise, radial resolution, absorbance scanning speed, and superior absorbance wavelength selection and detection. These instruments can collect high quality data fast enough to load up a full 8-hole rotor and collect data simultaneously at multiple wavelengths. This has revolutionized our virus analysis capabilities, allowing us to identify and quantify various viral fill states

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The KBI Biopharma Analytics Portal offers a one-stop-shop for quote requests, ordering, sample submission and shipment tracking, and a single visualization of studies. This fast and easy self-service process streamlines analytics ordering and sample coordination. 

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Explore Recent Resources


Adeno-Associated Virus-Mediated Gene Therapy: Analytical Advancements in Characterizing Size Distribution using SV-AUC and Multi-Wavelength Analysis for Accurate Identification of Resolved Species

At KBI Biopharma, we are leading the way in fully characterizing the size distribution of AAV products using SV-AUC combined with multi-wavelength analysis, giving us the ability to accurately identify each resolved species. 


Characterization and Quantification of Adeno-associated Virus Fill States Using Analytical Ultracentrifugation

In this poster, we show how this analysis can be used to identify empty, partially-packaged and full capsids, calculate the ratio of protein to DNA present in well resolved species, and compare the multi-wavelength results with expected values that arise from a consideration of the shape properties of AAVs. Furthermore, we show how this approach can help eliminate false peak identification. 


Comprehensive Size Distribution Analysis of Adeno-Associated Virus Fill States

In this poster, we compare the results from SV-AUC with MWA with two orthogonal separation methods: sedimentation equilibrium (SE) CsCl density gradient analysis and size exclusion chromatography coupled with multiple angle light scattering (SEC-MALS).

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