Analytical band centrifugation

Analytical band centrifugation (ABC) (also known as analytical band ultracentrifugation, or band sedimentation-velocity), is a specialized ultracentrifugation procedure, where unlike the typical use of (boundary) sedimentation velocity analytical ultracentrifugation (SV-AUC) wherein a homogenous bulk solution is centrifuged, in ABC a thin (~15 µL, ~500 µm)[1] sample is layered on top of a bulk solvent and then centrifuged. The method is distinguished from zone-sedimentation in that a stabilizing density gradient is self-generated during centrifugation, through the use of a higher density (than the sample) bulk "binary solvent", containing both a solvent (i.e. H2O), and a second component (small molecules, i.e. CsCl) that will sediment to form a stabilizing density gradient for the sample.[2]

Figure comparing an idealized time series from band ultracentrifugation and boundary ultracentrifugation experiments.

ABC also requires specially designed analytical ultracentrifuge cells, as the sample is not manually applied by pipette but instead automatically delivered via capillary under low g-forces at the beginning of a run from a reservoir within the cell. It was first demonstrated in 1963,[2] and was not commonly used for many decades,[3][4] but recently has become more widely used due to its applicability to quality control measurements on therapeutic viruses such as adeno-associated viruses (AAVs).[1] The profiles resulting from ABC analyses are similar in their interpretation to the profiles from electrophoretic separations ("electropherograms"), and thus have been dubbed "centrifugrams".[1]

See also

References

  1. Khasa, Harshit; Kilby, Greg; Chen, Xiaoyu; Wang, Chunlei (2021). "Analytical band centrifugation for the separation and quantification of empty and full AAV particles". Molecular Therapy - Methods & Clinical Development. Elsevier BV. 21: 585–591. doi:10.1016/j.omtm.2021.04.008. ISSN 2329-0501. PMC 8142049.
  2. Vinograd, Jerome; Bruner, Robert; Kent, Rebecca; Weigle, Jean (1963). "BAND-CENTRIFUGATION OF MACROMOLECULES AND VIRUSES IN SELF-GENERATING DENSITY GRADIENTS". Proceedings of the National Academy of Sciences. Proceedings of the National Academy of Sciences. 49 (6): 902–910. doi:10.1073/pnas.49.6.902. ISSN 0027-8424. PMC 300030.
  3. Schneider, Cornelia M.; Cölfen, Helmut (2018-06-21). "Analytical band centrifugation revisited". European Biophysics Journal. Springer Science and Business Media LLC. 47 (7): 799–807. doi:10.1007/s00249-018-1315-1. ISSN 0175-7571.
  4. Schneider, Cornelia M.; Haffke, Dirk; Cölfen, Helmut (2018-08-30). "Band Sedimentation Experiment in Analytical Ultracentrifugation Revisited". Analytical Chemistry. American Chemical Society (ACS). 90 (18): 10659–10663. doi:10.1021/acs.analchem.8b02768. ISSN 0003-2700.
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