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
- ^ a b c 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. 21. Elsevier BV: 585–591. doi: 10.1016/j.omtm.2021.04.008. ISSN 2329-0501. PMC 8142049. PMID 34095342.
- ^ a b 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. 49 (6): 902–910. doi: 10.1073/pnas.49.6.902. ISSN 0027-8424. PMC 300030. PMID 13997382.
- ^ Schneider, Cornelia M.; Cölfen, Helmut (2018-06-21). "Analytical band centrifugation revisited". European Biophysics Journal. 47 (7). Springer Science and Business Media LLC: 799–807. doi: 10.1007/s00249-018-1315-1. ISSN 0175-7571. PMID 29931388.
- ^ Schneider, Cornelia M.; Haffke, Dirk; Cölfen, Helmut (2018-08-30). "Band Sedimentation Experiment in Analytical Ultracentrifugation Revisited". Analytical Chemistry. 90 (18). American Chemical Society (ACS): 10659–10663. doi: 10.1021/acs.analchem.8b02768. ISSN 0003-2700. PMID 30160111.
.svg){kind=small} | This biochemistry article is a stub. You can help Wikipedia by expanding it. |
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
- ^ a b c 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. 21. Elsevier BV: 585–591. doi: 10.1016/j.omtm.2021.04.008. ISSN 2329-0501. PMC 8142049. PMID 34095342.
- ^ a b 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. 49 (6): 902–910. doi: 10.1073/pnas.49.6.902. ISSN 0027-8424. PMC 300030. PMID 13997382.
- ^ Schneider, Cornelia M.; Cölfen, Helmut (2018-06-21). "Analytical band centrifugation revisited". European Biophysics Journal. 47 (7). Springer Science and Business Media LLC: 799–807. doi: 10.1007/s00249-018-1315-1. ISSN 0175-7571. PMID 29931388.
- ^ Schneider, Cornelia M.; Haffke, Dirk; Cölfen, Helmut (2018-08-30). "Band Sedimentation Experiment in Analytical Ultracentrifugation Revisited". Analytical Chemistry. 90 (18). American Chemical Society (ACS): 10659–10663. doi: 10.1021/acs.analchem.8b02768. ISSN 0003-2700. PMID 30160111.
|
| This biochemistry article is a stub. You can help Wikipedia by expanding it. |