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

Huaying Zhao 1 Rodolfo Ghirlando Carlos Alfonso Fumio Arisaka Ilan Attali David L Bain 2 Marina M Bakhtina Donald F Becker 3 Gregory J Bedwell Ahmet Bekdemir Tabot M D Besong Catherine Birck 4 Chad A Brautigam William Brennerman Olwyn Byron Agnieszka Bzowska Jonathan B Chaires Catherine T Chaton 5 Helmut Cölfen 6 Keith D Connaghan Kimberly A Crowley Ute Curth Tina Daviter William L Dean Ana I Díez 7 Christine Ebel 8 Debra M Eckert 9 Leslie E Eisele Edward Eisenstein Patrick England 10 Carlos Escalante Jeffrey A Fagan Robert Fairman Ron M Finn Wolfgang Fischle 11 José García de la Torre Jayesh Gor Henning Gustafsson Damien Hall 12 Stephen E Harding 13 José G Hernández Cifre Andrew B Herr Elizabeth E Howell Richard S Isaac 14 Shu-Chuan Jao Davis Jose 15 Soon-Jong Kim Bashkim Kokona Jack A Kornblatt Dalibor Kosek Elena Krayukhina Daniel Krzizike Eric A Kusznir Hyewon Kwon 16 Adam Larson 17 Thomas M Laue Aline Le Roy 8 Andrew P Leech Hauke Lilie Karolin Luger Juan R Luque-Ortega Jia Ma 18 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 19 Jin-Ku Park 20 Peter D Pawelek Erby E Perdue Stephen J Perkins 21 Matthew A Perugini Craig L Peterson Martin G Peverelli Grzegorz Piszczek Gali Prag Peter E Prevelige Bertrand D E Raynal 10, 22 Lenka Rezabkova Klaus Richter 23 Alison E Ringel Rose Rosenberg 24 Arthur J Rowe 25 Arne C Rufer 26 David J Scott 27 Javier G Seravalli Alexandra S Solovyova 28 Renjie Song 29 David Staunton Caitlin Stoddard Katherine Stott Holger M Strauss 30 Werner W Streicher John P Sumida Sarah G Swygert Roman H Szczepanowski Ingrid Tessmer Ronald T Toth 31 Ashutosh Tripathy 32 Susumu Uchiyama Stephan F W Uebel Satoru Unzai Anna Vitlin Gruber 33 Peter H von Hippel Christine Wandrey Szu-Huan Wang 34 Steven E Weitzel Beata Wielgus-Kutrowska Cynthia Wolberger Martin Wolff Edward Wright 35 Yu-Sung Wu 36 Jacinta M Wubben Peter Schuck 37
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.
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Soumis le : mardi 15 septembre 2015 - 09:13:07
Dernière modification le : vendredi 5 février 2021 - 03:57:33

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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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