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

Langowski, J?rg; 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, Olwyn; Bzowska, Agnieszka; Chaires, Jonathan B.; Chaton, Catherine T.; C?lfen, Helmut; Connaghan, Keith D.; Crowley, Kimberly A.; Curth, Ute; Daviter, Tina; Dean, William L.; D?ez, Ana I.; Ebel, Christine; Eckert, Debra M.; Eisele, Leslie E.; Eisenstein, Edward; England, Patrick; Escalante, Carlos; Fagan, Jeffrey A.; Fairman, Robert; Finn, Ron M.; Fischle, Wolfgang; de la Torre, Jos? Garc?a; Gor, Jayesh; Gustafsson, Henning; Hall, Damien; Harding, Stephen E.; Cifre, Jos? G. Hern?ndez; Herr, Andrew B.; Howell, Elizabeth 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...



Huaying Zhao

Rodolfo Ghirlando

Carlos Alfonso

Fumio Arisaka

Ilan Attali

David L. Bain

Marina M. Bakhtina

Donald F. Becker

Gregory J. Bedwell

Ahmet Bekdemir

Tabot M.D. Besong

Catherine Birck

Chad A. Brautigam

William Brennerman

Olwyn Byron

Agnieszka Bzowska

Jonathan B. Chaires

Catherine T. Chaton


Keith D. Connaghan

Kimberly A. Crowley

Ute Curth

Tina Daviter

William L. Dean

Ana I.

Christine Ebel

Debra M. Eckert

Leslie E. Eisele

Edward Eisenstein

Patrick England

Carlos Escalante

Jeffrey A. Fagan

Robert Fairman

Ron M. Finn

Wolfgang Fischle

de la Torre

Jayesh Gor

Henning Gustafsson

Damien Hall


Andrew B. Herr

Elizabeth E. Howell

Richard S. Isaac

Shu-Chuan Jao

Davis Jose

Soon-Jong Kim

Bashkim Kokona

Jack A. Kornblatt

Dalibor Kosek

Elena Krayukhina

Daniel Krzizike

Eric A. Kusznir

Hyewon Kwon

Adam Larson

Thomas M. Laue

Aline Le Roy

Andrew P. Leech

Hauke Lilie

Karolin Luger

Juan R. Luque-Ortega

Jia Ma

Carrie A. May

Ernest L. Maynard

Anna Modrak-Wojcik

Yee-Foong Mok


Luitgard Nagel-Steger

Geeta J. Narlikar

Masanori Noda

Amanda Nourse

Tomas Obsil

Chad K. Park

Jin-Ku Park

Peter D. Pawelek

Erby E. Perdue

Stephen J. Perkins

Matthew A. Perugini

Craig L. Peterson

Martin G. Peverelli

Grzegorz Piszczek

Gali Prag

Peter E. Prevelige

Bertrand D.E. Raynal

Lenka Rezabkova

Klaus Richter

Alison E. Ringel

Rose Rosenberg

Arthur J. Rowe

Arne C. Rufer

Associate Professor & Reader in Physical Biochemistry

Javier G. Seravalli

Alexandra S. Solovyova

Renjie Song

David Staunton

Caitlin Stoddard

Katherine Stott

Holger M. Strauss

Werner W. Streicher

John P. Sumida

Sarah G. Swygert

Roman H. Szczepanowski

Ingrid Tessmer

Ronald T. Toth

Ashutosh Tripathy

Susumu Uchiyama

Stephan F.W. Uebel

Satoru Unzai

Anna Vitlin Gruber

Peter H. von Hippel

Christine Wandrey

Szu-Huan Wang

Steven E. Weitzel

Beata Wielgus-Kutrowska

Cynthia Wolberger

Martin Wolff

Edward Wright

Yu-Sung Wu

Jacinta M. Wubben

Peter Schuck




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.


Langowski, J., Zhao, H., Ghirlando, R., Alfonso, C., Arisaka, F., Attali, I., …Schuck, P. (2015). A Multilaboratory Comparison of Calibration Accuracy and the Performance of External References in Analytical Ultracentrifugation. PLoS ONE, 10(5), Article e0126420.

Journal Article Type Article
Acceptance Date Apr 2, 2015
Online Publication Date May 21, 2015
Publication Date May 21, 2015
Deposit Date Apr 3, 2017
Publicly Available Date Apr 3, 2017
Journal PLoS ONE
Electronic ISSN 1932-6203
Publisher Public Library of Science
Peer Reviewed Peer Reviewed
Volume 10
Issue 5
Article Number e0126420
Public URL
Publisher URL


journal.pone.0126420 (1).pdf (1.8 Mb)

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