Missing aggregates in SEC-MALS results

Questions about Multi-Angle Light Scattering coupled with Size-Exclusion Chromatography or Field-Flow Fractionation

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If higher order aggregates are retaining in the column frit or not eluting from the column, how do you say that SEC-MALS results are accurate?
SEC-MALS (as with any other technique) can only analyze what is actually present in the sample, so if higher-order aggregates are filtered out or are retained on the column, we will miss part of the sample. In addition, the MALS signal is an average of all species that are passing through the detector at a particular instant in time, so the purer the eluting peaks, the more accurate our molecular weight calculation will be. All of this means we must choose chromatography conditions that are appropriate for the sample to 1) analyze and fractionate all species present in solution; 2) avoid column interactions; and 3) maximize peak resolution.

[answer provided by Mark Spears, Ph.D., Fujifilm Diosynth Biotechnologies]
Daniel Some, Ph.D.
Principal Scientist
Wyatt Technology Corp.
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Aggregates that are too large to elute from the column or are condensed in the void volume may be characterized by a related method, FFF-MALS. FFF uses an open channel to separate macromolecules and particles by size, and does not block, shear or retain large aggregates.

For more information on FFF-MALS, click on http://bit.ly/2VruTPb
Daniel Some, Ph.D.
Principal Scientist
Wyatt Technology Corp.
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