Limiting the sedimentation coefficient for sedimentation velocity data analysis: Partial boundary modeling and g( s ∗) approaches revisited

Brown and coworkers (Eur. Biophys. J. 38 (2009) 1079–1099) introduced partial boundary modeling (PBM) to simplify sedimentation velocity data analysis by excluding species outside the range of interest (e.g., aggregates, impurities) via restricting the sedimentation coefficient range being fitted. T...

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Veröffentlicht in:Analytical biochemistry 2011-05, Vol.412 (2), p.189-202
1. Verfasser: Philo, John S.
Format: Artikel
Sprache:eng
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Zusammenfassung:Brown and coworkers (Eur. Biophys. J. 38 (2009) 1079–1099) introduced partial boundary modeling (PBM) to simplify sedimentation velocity data analysis by excluding species outside the range of interest (e.g., aggregates, impurities) via restricting the sedimentation coefficient range being fitted. They strongly criticized the alternate approach of fitting g( s ∗) distributions using similar range limits, arguing that (i) it produces “nonoptimal fits in the original data space” and (ii) the g( s ∗) data transformations lead to gross underestimates of the parameter confidence intervals. It is shown here that neither of those criticisms is valid. These two approaches are not truly fitting the same data or in equivalent ways; thus, they should not actually give the same best-fit parameters. The confidence limits for g( s ∗) fits derived using F statistics, bootstrap, or a new Monte Carlo algorithm are in good agreement and show no evidence for significant statistical distortion. Here 15 g( s ∗) measurements on monoclonal antibody samples gave monomer mass estimates with experimental standard deviations of less than 1%, close to the confidence limit estimates. Tests on both real and simulated data help to clarify the strengths and drawbacks of both approaches. New algorithms for computing g( s ∗) and a scan-differencing approach for PBM are introduced.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2011.01.035