A Multilaboratory Comparison of Calibration Accuracy and the Performance of External References in Analytical Ultracentrifugation

摘要: Analytical ultracentrifugation (AUC) is a first principles based method to determine absolute sedimentation coefficients and buoyant molar masses of macromolecules their complexes, reporting on size shape in free solution. The purpose this multi-laboratory study was establish the precision accuracy basic data dimensions AUC validate previously proposed calibration techniques. Three kits cell assemblies containing radial temperature tools bovine serum albumin (BSA) reference sample were shared among 67 laboratories, generating 129 comprehensive sets. These allowed for an assessment many parameters instrument performance, including reported scan time after start centrifugation, calibration, magnification. range obtained BSA monomer different instruments using optical systems from 3.655 S 4.949 S, with mean standard deviation (4.304 ± 0.188) (4.4%). After combined application correction factors derived external references elapsed time, velocity, temperature, magnification, s-values reduced 7-fold 4.325 6-fold 0.030 (0.7%). In addition, large set provided opportunity instrument-to-instrument variation positions files, photometric or refractometric signal magnitudes, calculated apparent mass fraction dimers. results highlight necessity effectiveness independent reliable quantitative studies.