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

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]

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. 1 2 3 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. 1 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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