TY - JOUR
T1 - Recorded scan times can limit the accuracy of sedimentation coefficients in analytical ultracentrifugation
AU - Zhao, Huaying
AU - Ghirlando, Rodolfo
AU - Piszczek, Grzegorz
AU - Curth, Ute
AU - Brautigam, Chad A.
AU - Schuck, Peter
N1 - Funding Information:
This work was supported by the Intramural Research Programs of the National Institute of Biomedical Imaging and Bioengineering, National Institute of Diabetes and Digestive and Kidney Diseases, and National Heart, Lung, and Blood Institute, National Institutes of Health .
PY - 2013/6/1
Y1 - 2013/6/1
N2 - We report systematic and large inaccuracies in the recorded elapsed time in data files from the analytical ultracentrifuge, leading to overestimates of the sedimentation coefficients of up to 10%. This far exceeds previously considered factors contributing to the uncertainty in this parameter and has significant ramifications for derived parameters such as hydrodynamic shape and molar mass estimates. The source of this error is currently unknown, but we found it to be quantitatively consistent across different instruments, increasing with rotor speed. Furthermore, its occurrence appears to correlate with the use of the latest data acquisition software from the manufacturer, in use in some of our laboratories for nearly 2 years. Many of the recently published sedimentation coefficients may need to be reexamined. The problem can be easily recognized by comparing the file timestamps provided by the operating system with the elapsed scan times recorded within the data files. Therefore, we implemented a routine in SEDFIT that can automatically examine the data files, alert the user to significant discrepancies, and correct the scan times accordingly. This eliminates errors in the recorded scan times.
AB - We report systematic and large inaccuracies in the recorded elapsed time in data files from the analytical ultracentrifuge, leading to overestimates of the sedimentation coefficients of up to 10%. This far exceeds previously considered factors contributing to the uncertainty in this parameter and has significant ramifications for derived parameters such as hydrodynamic shape and molar mass estimates. The source of this error is currently unknown, but we found it to be quantitatively consistent across different instruments, increasing with rotor speed. Furthermore, its occurrence appears to correlate with the use of the latest data acquisition software from the manufacturer, in use in some of our laboratories for nearly 2 years. Many of the recently published sedimentation coefficients may need to be reexamined. The problem can be easily recognized by comparing the file timestamps provided by the operating system with the elapsed scan times recorded within the data files. Therefore, we implemented a routine in SEDFIT that can automatically examine the data files, alert the user to significant discrepancies, and correct the scan times accordingly. This eliminates errors in the recorded scan times.
KW - Hydrodynamic modeling
KW - Sedimentation velocity
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U2 - 10.1016/j.ab.2013.02.011
DO - 10.1016/j.ab.2013.02.011
M3 - Article
C2 - 23458356
AN - SCOPUS:84876134886
SN - 0003-2697
VL - 437
SP - 104
EP - 108
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -