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

Zhao, H., Ghirlando, R., Alfonso, C., Arisaka, F., Attali, H., Bain, D. L., et al. (2015). A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation. PLOS One, 10(5): e0126420. doi:10.1371/journal.pone.0126420.

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Zhao, H., Author
Ghirlando, R., Author
Alfonso, C., Author
Arisaka, F., Author
Attali, H., Author
Bain, D. L., Author
Bakhtina, M. M., Author
Becker, D. F., Author
Bedwell, G. J., Author
Bekdemir, A., Author
Besong, T. M. D., Author
Birck, C., Author
Brautigam, C. A., Author
Brennerman, W., Author
Byron, O., Author
Bzowska, A., Author
Chaires, J. B., Author
Chaton, C. T., Author
Cölfen, H., Author
Connaghan, K. D., Author
Crowley, K. A., AuthorCurth, U., AuthorDaviter, T., AuthorDean, W. L., AuthorDíez, A. I., AuthorEbel, C., AuthorEckert, D. M., AuthorEisele, L. E., AuthorEisenstein, E., AuthorEngland, P., AuthorEscalante, C., AuthorFagan, J. A., AuthorFairman, R., AuthorFinn, R. M., AuthorFischle, W.1, Author              García de la Torre, J., AuthorGor, J., AuthorGustafsson, H., AuthorHall, D., AuthorHarding, S. E., AuthorHernández Cifre, J. G. , AuthorHerr, A. B., AuthorHowell, E. E., AuthorIsaac, R. S., AuthorJao, S. C., AuthorJose, Davis, AuthorKim, S. J., AuthorKokona, B., AuthorKornblatt, J. A., AuthorKosek, D., AuthorKrayukhinac, E., AuthorKrzizike, D., AuthorKusznir, E. A., AuthorKwon, H., AuthorLarson, A., AuthorLaue, T. M., AuthorLe Roy, A., AuthorLeech, A. P., AuthorLilie, H., AuthorLuger, K., AuthorLuque-Ortega, J. R., AuthorMa, J., AuthorMay, C. A., AuthorMaynard, E. L., AuthorModrak-Wojcik, A., AuthorMok, Y. F., AuthorMücke, N., AuthorNagel-Steger, L., AuthorNarlikar, G. J., AuthorNodac, M., AuthorNourse, A., AuthorObsil, T., AuthorPark, C. K., AuthorPark, J. K., AuthorPawelek, P. D., AuthorPerdue, E. E., AuthorPerkins, S. J., AuthorPerugini, M. A., AuthorPeterson, C. L., AuthorPeverelli, M. G., AuthorPiszczek, G., AuthorPrag, G., AuthorPrevelige, P. E., AuthorRaynal, B. D. E., AuthorRezabkova, L., AuthorRichter, K., AuthorRingel, A. E., AuthorRosenberg, R., AuthorRowe, A. J., AuthorRufer, A. C., AuthorScott, D. J., AuthorSeravalli, J. G., AuthorSolovyova, A. S., AuthorSong, R., AuthorStaunton, D., AuthorStoddard, C., AuthorStott, K., AuthorStrauss, H. M., AuthorStreicherd, W. W., AuthorSumida, J. P., AuthorSwygert, S. G., AuthorSzczepanowski, R. H., AuthorTessmer, I., AuthorToth, R. T., AuthorTripathy, A., AuthorUchiyama, S., AuthorUebel, S. F. W., AuthorUnzai, S., AuthorVitlin, A., AuthorGruber, P., Authorvon Hippel, H., AuthorWandrey, C., AuthorWang, Szu-Huan, AuthorWeitzel, S. E., AuthorWielgus-Kutrowska, B., AuthorWolberger, C., AuthorWolff, M., AuthorWright, E., AuthorWu, Y. S., AuthorWubben, J. M., AuthorSchuck, P., Author more..
1Research Group of Chromatin Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578604              


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 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.


Language(s): eng - English
 Dates: 2015-05-21
 Publication Status: Published online
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1371/journal.pone.0126420
 Degree: -



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Title: PLOS One
Source Genre: Journal
Publ. Info: -
Pages: 30 Volume / Issue: 10 (5) Sequence Number: e0126420 Start / End Page: - Identifier: -