A new UltraScan module for the characterization and quantification of analytical buoyant density equilibrium experiments to determine AAV capsid loading
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Date
2023
Authors
Savelyev, Alexey
Brookes, Emre H.
Henrickson, Amy
Demeler, Borries
Journal Title
Journal ISSN
Volume Title
Publisher
Springer
Abstract
A method for characterizing and quantifying peaks formed in an analytical buoyant density
equilibrium (ABDE) experiment is presented. An algorithm is derived to calculate the
concentration of the density forming gradient material at every point in the cell, provided the rotor
speed, temperature, meniscus position, bottom of the cell position, and the loading concentration,
molar mass, and partial specific volume of the density gradient-forming material are known. In
addition, a new peak fitting algorithm has been developed which allows the user to automatically
quantify the peaks formed in terms of density, apparent partial specific volume, and relative
abundance. The method is suitable for both ionic and non-ionic density forming materials and
can be used with data generated from the UV optical system as well as the AVIV fluorescence
optical system. These methods have been programmed in a new UltraScan-III module (us_abde).
Examples are shown that demonstrate the application of the new module to adeno-associated viral
vector preparations and proteins.
Description
Accepted author manuscript
Keywords
Analytical ultracentrifugation , Analytical buoyant density equilibrium , AAV quantification and characterization , Peak fitting , UltraScan software
Citation
Savelyev, A., Brookes, E. H., Henrickson, A., & Bemeler, B. (2023). A new UltraScan module for the characterization and quantification of analytical buoyant density equilibrium experiments to determine AAV capsid loading. European Biophysics Journal, 52(4-5), 311-320. https://doi.org/10.1007/s00249-023-01641-4