Analytical ultracentrifugation

Analytical ultracentrifugation is an analytical technique which combines an ultracentrifuge with optical monitoring systems.

Spinco Model E Analytical Ultracentrifuge. This is an early instrument, dating to the 1950s. The operator is seated in front of the sample chamber, with his left hand touching the rotor. In operation, the chamber would be sealed behind an armored shroud and pumped down to vacuum. The shroud has been lowered to allow access for loading or unloading the rotor.[1]

In an analytical ultracentrifuge (commonly abbreviated as AUC), a sample’s sedimentation profile is monitored in real time by an optical detection system. The sample is detected via ultraviolet light absorption and/or interference optical refractive index sensitive system, monitored by light-sensitive diode array or by film in the older machines. The operator can thus observe the change of sample concentration versus the axis of the rotation profile with time as a result of the applied centrifugal field. With modern instrumentation, these observations are electronically digitized and stored for further mathematical analysis.

The information that can be obtained from an analytical ultracentrifuge includes the gross shape of macromolecules, conformational changes in macromolecules, and size distributions of macromolecules. With AUC it is possible to gain information on the number and subunit stoichiometry of non-covalent complexes and equilibrium constants of macromolecules such as proteins, DNA, nanoparticles or other assemblies from different molecule classes. The simplest measurement to be obtained is the sedimentation coefficient, which depends upon the size of the molecules being sedimented. This is the ratio of a particle's sedimentation velocity to the applied acceleration causing the sedimentation.

Analytical ultracentrifugation has recently seen a rise in use because of increased ease of analysis with modern computers and the development of software, including a National Institutes of Health supported software package, SedFit.

  1. ^ "Technical Manual, Spinco Ultracentrifuge Model E". Science History Institute Digital Collections. Retrieved 2018-12-18.