A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation
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...
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| Format: | Article |
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Public Library of Science
2015
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| Online Access: | https://eprints.nottingham.ac.uk/41716/ |
| _version_ | 1848796338636980224 |
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| author | Langowski, Jörg 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. Cölfen, Helmut Connaghan, Keith D. Crowley, Kimberly A. Curth, Ute Daviter, Tina Dean, William L. Díez, 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, José García Gor, Jayesh Gustafsson, Henning Hall, Damien Harding, Stephen E. Cifre, José G. Hernández 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, 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 Mücke, 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. Tripathy, Ashutosh Uchiyama, Susumu Uebel, Stephan F.W. Unzai, Satoru Gruber, Anna Vitlin von Hippel, Peter H. Wandrey, Christine Wang, Szu-Huan Weitzel, Steven E. Wielgus-Kutrowska, Beata Wolberger, Cynthia Wolff, Martin Wright, Edward Wu, Yu-Sung Wubben, Jacinta M. Schuck, Peter |
| author_facet | Langowski, Jörg 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. Cölfen, Helmut Connaghan, Keith D. Crowley, Kimberly A. Curth, Ute Daviter, Tina Dean, William L. Díez, 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, José García Gor, Jayesh Gustafsson, Henning Hall, Damien Harding, Stephen E. Cifre, José G. Hernández 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, 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 Mücke, 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. Tripathy, Ashutosh Uchiyama, Susumu Uebel, Stephan F.W. Unzai, Satoru Gruber, Anna Vitlin von Hippel, Peter H. Wandrey, Christine Wang, Szu-Huan Weitzel, Steven E. Wielgus-Kutrowska, Beata Wolberger, Cynthia Wolff, Martin Wright, Edward Wu, Yu-Sung Wubben, Jacinta M. Schuck, Peter |
| author_sort | Langowski, Jörg |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:46:24Z |
| format | Article |
| id | nottingham-41716 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:46:24Z |
| publishDate | 2015 |
| publisher | Public Library of Science |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-417162020-05-04T17:08:16Z https://eprints.nottingham.ac.uk/41716/ A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation Langowski, Jörg 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. Cölfen, Helmut Connaghan, Keith D. Crowley, Kimberly A. Curth, Ute Daviter, Tina Dean, William L. Díez, 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, José García Gor, Jayesh Gustafsson, Henning Hall, Damien Harding, Stephen E. Cifre, José G. Hernández 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, 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 Mücke, 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. Tripathy, Ashutosh Uchiyama, Susumu Uebel, Stephan F.W. Unzai, Satoru Gruber, Anna Vitlin von Hippel, Peter H. Wandrey, Christine Wang, Szu-Huan Weitzel, Steven E. Wielgus-Kutrowska, Beata Wolberger, Cynthia Wolff, Martin Wright, Edward Wu, Yu-Sung Wubben, Jacinta M. Schuck, Peter 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. Public Library of Science 2015-05-21 Article PeerReviewed Langowski, Jörg, 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., Cölfen, Helmut, Connaghan, Keith D., Crowley, Kimberly A., Curth, Ute, Daviter, Tina, Dean, William L., Díez, 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, José García, Gor, Jayesh, Gustafsson, Henning, Hall, Damien, Harding, Stephen E., Cifre, José G. Hernández, 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, 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, Mücke, 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., Tripathy, Ashutosh, Uchiyama, Susumu, Uebel, Stephan F.W., Unzai, Satoru, Gruber, Anna Vitlin, von Hippel, Peter H., Wandrey, Christine, Wang, Szu-Huan, Weitzel, Steven E., Wielgus-Kutrowska, Beata, Wolberger, Cynthia, Wolff, Martin, Wright, Edward, Wu, Yu-Sung, Wubben, Jacinta M. and Schuck, Peter (2015) A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation. PLoS ONE, 10 (5). e0126420/1-e0126420/30. ISSN 1932-6203 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0126420 doi:10.1371/journal.pone.0126420 doi:10.1371/journal.pone.0126420 |
| spellingShingle | Langowski, Jörg 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. Cölfen, Helmut Connaghan, Keith D. Crowley, Kimberly A. Curth, Ute Daviter, Tina Dean, William L. Díez, 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, José García Gor, Jayesh Gustafsson, Henning Hall, Damien Harding, Stephen E. Cifre, José G. Hernández 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, 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 Mücke, 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. Tripathy, Ashutosh Uchiyama, Susumu Uebel, Stephan F.W. Unzai, Satoru Gruber, Anna Vitlin von Hippel, Peter H. Wandrey, Christine Wang, Szu-Huan Weitzel, Steven E. Wielgus-Kutrowska, Beata Wolberger, Cynthia Wolff, Martin Wright, Edward Wu, Yu-Sung Wubben, Jacinta M. Schuck, Peter A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation |
| title | A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation |
| title_full | A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation |
| title_fullStr | A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation |
| title_full_unstemmed | A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation |
| title_short | A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation |
| title_sort | multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation |
| url | https://eprints.nottingham.ac.uk/41716/ https://eprints.nottingham.ac.uk/41716/ https://eprints.nottingham.ac.uk/41716/ |