A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation.

Huaying Zhao 1 Rodolfo Ghirlando Carlos Alfonso 2 Fumio Arisaka Ilan Attali David L Bain 3 Marina M Bakhtina Donald F Becker 4 Gregory J Bedwell Ahmet Bekdemir Tabot M D Besong Catherine Birck 5 Chad A Brautigam William Brennerman Olwyn Byron Agnieszka Bzowska Jonathan B Chaires Catherine T Chaton 6 Helmut Cölfen 7 Keith D Connaghan Kimberly A Crowley Ute Curth Tina Daviter William L Dean Ana I Díez 8 Christine Ebel 9 Debra M Eckert 10 Leslie E Eisele Edward Eisenstein Patrick England 11 Carlos Escalante Jeffrey A Fagan Robert Fairman Ron M Finn Wolfgang Fischle 12 José García De La Torre Jayesh Gor Henning Gustafsson Damien Hall 13 Stephen E Harding 14 José G Hernández Cifre Andrew B Herr Elizabeth E Howell Richard S Isaac 15 Shu-Chuan Jao Davis Jose 16 Soon-Jong Kim Bashkim Kokona Jack A Kornblatt Dalibor Kosek Elena Krayukhina Daniel Krzizike Eric A Kusznir Hyewon Kwon 17 Adam Larson 18 Thomas M Laue Aline Le Roy 9 Andrew P Leech Hauke Lilie Karolin Luger Juan R Luque-Ortega Jia Ma 19 Carrie A May Ernest L Maynard Anna Modrak-Wojcik Yee-Foong Mok Norbert Mücke Luitgard Nagel-Steger Geeta J Narlikar Masanori Noda Amanda Nourse Tomas Obsil Chad K Park 20 Jin-Ku Park 21 Peter D Pawelek Erby E Perdue Stephen J Perkins 22 Matthew A Perugini Craig L Peterson Martin G Peverelli Grzegorz Piszczek Gali Prag Peter E Prevelige Bertrand D E Raynal 11, 23 Lenka Rezabkova Klaus Richter 24 Alison E Ringel Rose Rosenberg 25 Arthur J Rowe 26 Arne C Rufer 27 David J Scott 28 Javier G Seravalli Alexandra S Solovyova 29 Renjie Song 30 David Staunton Caitlin Stoddard Katherine Stott Holger M Strauss 31 Werner W Streicher John P Sumida Sarah G Swygert Roman H Szczepanowski Ingrid Tessmer Ronald T Toth 32 Ashutosh Tripathy 33 Susumu Uchiyama Stephan F W Uebel Satoru Unzai Anna Vitlin Gruber 34 Peter H Von Hippel Christine Wandrey Szu-Huan Wang 35 Steven E Weitzel Beata Wielgus-Kutrowska Cynthia Wolberger Martin Wolff Edward Wright 36 Yu-Sung Wu 37 Jacinta M Wubben Peter Schuck 38
16 DCM - CIRE - Département de Chimie Moléculaire - Chimie Inorganique Redox Biomimétique
CNRS - Centre National de la Recherche Scientifique : UMR5250, UJF - Université Joseph Fourier - Grenoble 1, Institut de Chimie Moléculaire de Grenoble (ICMG)
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.
Type de document :
Article dans une revue
PLoS ONE, Public Library of Science, 2015, 10 (5), pp.e0126420. 〈10.1371/journal.pone.0126420〉
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Huaying Zhao, Rodolfo Ghirlando, Carlos Alfonso, Fumio Arisaka, Ilan Attali, et al.. A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation.. PLoS ONE, Public Library of Science, 2015, 10 (5), pp.e0126420. 〈10.1371/journal.pone.0126420〉. 〈hal-01199168〉



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