A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation
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Zhao, Huaying; Ghirlando, Rodolfo; Alfonso, Carlos; Arisaka, Fumio; Attali, Ilan; Bain, David L.; Bakhtina, Marina M.; Becker, Donald F.; Bedwell, Gregory J.; Bekdemir, Ahmet; Hall, Damien
Description
Analytical ultracentrifugation (AUC) is a first principles based method to determine absolute sedimentation coefficients and buoyant molar masses of macromolecules and their complexes, reporting on their size and shape in free solution. The purpose of this multi-laboratory study was to establish the precision and accuracy of basic data dimensions in AUC and validate previously proposed calibration techniques. Three kits of AUC cell assemblies containing radial and temperature calibration tools...[Show more]
dc.contributor.author | Zhao, Huaying | |
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dc.contributor.author | Ghirlando, Rodolfo | |
dc.contributor.author | Alfonso, Carlos | |
dc.contributor.author | Arisaka, Fumio | |
dc.contributor.author | Attali, Ilan | |
dc.contributor.author | Bain, David L. | |
dc.contributor.author | Bakhtina, Marina M. | |
dc.contributor.author | Becker, Donald F. | |
dc.contributor.author | Bedwell, Gregory J. | |
dc.contributor.author | Bekdemir, Ahmet | |
dc.contributor.author | Hall, Damien | |
dc.date.accessioned | 2015-12-13T22:15:36Z | |
dc.identifier.issn | 1932-6203 | |
dc.identifier.uri | http://hdl.handle.net/1885/70478 | |
dc.description.abstract | Analytical ultracentrifugation (AUC) is a first principles based method to determine absolute sedimentation coefficients and buoyant molar masses of macromolecules and their complexes, reporting on their size and shape in free solution. The purpose of this multi-laboratory study was to establish the precision and accuracy of basic data dimensions in AUC and validate previously proposed calibration techniques. Three kits of AUC cell assemblies containing radial and temperature calibration tools and a bovine serum albumin (BSA) reference sample were shared among 67 laboratories, generating 129 comprehensive data sets. These allowed for an assessment of many parameters of instrument performance, including accuracy of the reported scan time after the start of centrifugation, the accuracy of the temperature calibration, and the accuracy of the radial magnification. The range of sedimentation coefficients obtained for BSA monomer in different instruments and using different optical systems was from 3.655 S to 4.949 S, with a mean and standard deviation of (4.304 ± 0.188) S (4.4%). After the combined application of correction factors derived from the external calibration references for elapsed time, scan velocity, temperature, and radial magnification, the range of s-values was reduced 7-fold with a mean of 4.325 S and a 6-fold reduced standard deviation of ± 0.030 S (0.7%). In addition, the large data set provided an opportunity to determine the instrument-to-instrument variation of the absolute radial positions reported in the scan files, the precision of photometric or refractometric signal magnitudes, and the precision of the calculated apparent molar mass of BSA monomer and the fraction of BSA dimers. These results highlight the necessity and effectiveness of independent calibration of basic AUC data dimensions for reliable quantitative studies. | |
dc.publisher | Public Library of Science | |
dc.rights | Author/s retain copyright | |
dc.source | PLOS ONE (Public Library of Science) | |
dc.title | A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation | |
dc.type | Journal article | |
local.description.notes | Imported from ARIES | |
local.identifier.citationvolume | 10 | |
dc.date.issued | 2015 | |
local.identifier.absfor | 039904 - Organometallic Chemistry | |
local.identifier.absfor | 030503 - Organic Chemical Synthesis | |
local.identifier.absfor | 030701 - Quantum Chemistry | |
local.identifier.ariespublication | a383154xPUB2324 | |
local.type.status | Published Version | |
local.contributor.affiliation | Zhao, Huaying, National Institute of Biomedical Imaging and Bioengineering | |
local.contributor.affiliation | Ghirlando, Rodolfo, National Institute of Diabetes and Digestive and Kidney Diseases | |
local.contributor.affiliation | Alfonso, Carlos, Centro de Investigaciones Biológicas | |
local.contributor.affiliation | Arisaka, Fumio, Nihon University | |
local.contributor.affiliation | Attali, Ilan, Tel Aviv University | |
local.contributor.affiliation | Bain, David L., University of Colorado Denver Anschutz Medical Campus | |
local.contributor.affiliation | Bakhtina, Marina M., The Ohio State University | |
local.contributor.affiliation | Becker, Donald F., University of Nebraska-Lincoln | |
local.contributor.affiliation | Bedwell, Gregory J., University of Alabama at Birmingham | |
local.contributor.affiliation | Bekdemir, Ahmet, École Polytechnique Fédérale de Lausanne | |
local.contributor.affiliation | Hall, Damien, College of Physical and Mathematical Sciences, ANU | |
local.bibliographicCitation.issue | 5 | |
local.bibliographicCitation.startpage | 1 | |
local.bibliographicCitation.lastpage | 30 | |
local.identifier.doi | 10.1371/journal.pone.0126420 | |
dc.date.updated | 2015-12-11T07:18:46Z | |
local.identifier.scopusID | 2-s2.0-84930615100 | |
dcterms.accessRights | Open Access | |
Collections | ANU Research Publications |
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