[RASMB] non-ideality in velocity [was: interference optics]
Ewa Folta-Stogniew
ef55 at email.med.yale.edu
Tue Apr 8 08:02:38 PDT 2008
there is not much published that directly compares SEC-MALLS to SV, but in
published study:
Gabrielson J P, Brader M. L., Pekar A. H., Mathis K. B., Winter G.,
Carpenter J. F., and Randolph T. W. (2007) Quantitation of aggregate levels
in a recombinant humanized monoclonal antibody formulation by
size-exclusion chromatography, asymmetrical flow field flow fractionation,
and sedimentation velocity. J Pharm.Sci 96: 268-279
>Incidentally, it has been my experience that SEC-MALS is not so great
>at detecting large irreversible aggregates, because they can get stuck in
>the SEC column and you never see them in the MALS detector. However,
>the reversible oligomerization may be detected fine by MALS .
seems not be the case.
My guess would be that the reversible are harder for SEC-MALLS to account
for due to a constant dilution on SEC and an introduction of another phase
with which the protein can interact (SEC matrix) that may disturb the
equilibrium.
Ewa
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