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

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Issue Date
2015-05-21Author
Zhao, Huaying
Ghirlando, Rodolfo
Alfonso, Carlos
Arisaka, Fumio
Attali, Ilan
Bain, David L.
Bakhtina, Marina M.
Becker, Donald F.
Bedwell, Gregory J.
Bekdemir, Ahmet
Besong, Tabot M. D.
Birck, Catherine
Brautigam, Chad A.
Brennerman, William
Byron, Owlyn
Bzowska, Agnieszka
Chaires, Johnathan B.
Chaton, Catherine T.
Coelfen, Helmut
Connaghan, Keith D.
Crowley, Kimberly A.
Curth, Ute
Daviter, Tina
Dean, William L.
Diez, Ana I.
Ebel, Christine
Eckert, Debra M.
Eisele, Leslie E.
Eisenstein, Edward
England, Patrick
Escalante, Carlos
Fagan, Jeffery A.
Fairman, Robert
Finn, Ron M.
Fischle, Wolfgang
de la Torre, Jose Garcia
Gor, Jayesh
Gustafsson, Henning
Hall, Damien
Harding, Stephen E.
Cifre, Jose G. Hernadez
Herr, Andrew B.
Howell, Elizebeth E.
Isaac, Richard S.
Jao, Shu-Chuan
Jose, Davis
Kim, Soon-Jong
Kokona, Bashkim
Kornblatt, Jack A.
Kosek, Dalibor
Krayukhina, Elena
Krzizike, Daniel
Kusznir, Eric A.
Kwon, Hyewon
Larson, Adam
Laue, Thomas M.
Le Roy, Aline
Leech, Andrew P.
Lilie, Hauke
Luger, Karolin
Luque-Ortega, Juan R.
Ma, Jia
May, Carrie A.
Maynard, Ernest L.
Modrak-Wojcik, Anna
Mok, Yee-Foong
Muecke, Norbert
Nagel-Steger, Luitgard
Narlikar, Geeta J.
Noda, Masanori
Nourse, Amanda
Obsil, Tomas
Park, Chad K.
Park, Jin-Ku
Pawelek, Peter D.
Perdue, Erby E.
Perkins, Stephen J.
Perugini, Matthew A.
Peterson, Craig L.
Peverelli, Martin G.
Piszczek, Grzegorz
Prag, Gali
Prevelige, Peter E.
Raynal, Bertrand D. E.
Rezabkova, Lenka
Richter, Klaus
Ringel, Alison E.
Rosenberg, Rose
Rowe, Arthur J.
Rufer, Arne C.
Scott, David J.
Seravalli, Javier G.
Solovyova, Alexandra S.
Song, Renjie
Staunton, David
Stoddard, Caitlin
Stott, Katherine
Strauss, Holger M.
Streicher, Werner W.
Sumida, John P.
Swygert, Sarah G.
Szczepanowski, Roman H.
Tessmer, Ingrid
Toth, Ronald T., IV
Tripathy, Ashutosh
Uchiyama, Susumu
Uebel, Stephan F. W.
Unazi, Satoru
Gruben, Anna Vitlin
von Hippel, Peter H.
Wandrey, Christine
Wang, Szu-Huan
Weitzel, Steven E.
Wielgu-Kutrowska, Beata
Wolberger, Cynthia
Wolff, Martin
Wright, Edward
Yu-Sung, Wu
Wubben, Jacinta M.
Schuck, Peter
Publisher
Public Library of Science
Type
Article
Article Version
Scholarly/refereed, publisher version
Rights
This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication
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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.
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Citation
Zhao H, Ghirlando R, Alfonso C, Arisaka F, Attali I, Bain DL, et al. (2015) A Multilaboratory Comparison of Calibration Accuracy and the Performance of External References in Analytical Ultracentrifugation. PLoS ONE 10(5): e0126420.
http://dx.doi.org/10.1371/journal.pone.0126420
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Except where otherwise noted, this item's license is described as: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication