[RASMB] analysis of biotech samples [was: non-ideality in SV]
Allen Minton
minton at helix.nih.gov
Tue Apr 8 10:17:52 PDT 2008
At 12:20 PM 4/8/2008, you wrote:
> (3) The regulatory agencies would indeed like
> to know the exact distribution of oligomers,
> both reversible and irreversible, in the
> product vial, even for products at 100+ mg/mL.
> Unfortunately they don't always understand that
> this may be technically impossible with current
> technology and current theoretical
> understanding of the strong non-ideality effects that may be present.
We've developed some approximate methods for
interpretation of static light scattering,
sedimentation equilibrium, and osmotic pressure
of highly concentrated (> 100 mg/ml) protein
solutions containing multiple macrosolute species
(including both reversibly and irreversibly
formed oligomers). We know the conditions under
which the approximations in the model ought to be
valid, and the methods seem to work pretty well
when applied to data obtained under those
conditions. Analyses of data obtained by
different methods from the same protein yield
internally consistent results. If you are
interested, check out the following publications:
Minton AP. 2008. Effective hard particle model
for the osmotic pressure of highly concentrated
binary protein solutions. Biophys. J 94: L57-59.
Jiménez M, Rivas G, Minton AP. 2007. Quantitative
characterization of weak self-association in
concentrated solutions of immunoglobulin G via
measurement of sedimentation equilibrium and
osmotic pressure. Biochemistry 45: 13356-60.
Minton AP. 2007. Static light scattering from
concentrated protein solutions. I. General
theory for protein mixtures and application to
self-associating proteins. Biophys J 93: 1321-8.
Minton AP. 2007. The effective hard particle
model provides a simple, robust and broadly
applicable description of nonideal behavior in
concentrated solutions of bovine serum albumin
and other non-associating proteins. J Pharm Sci 96: 3466-69.
Zorrilla S, Jimenez M, Lillo P, Rivas G, Minton
AP. 2004. Sedimentation equilibrium in a solution
containing an arbitrary number of solute species
at arbitrary concentrations: theory and
application to concentrated solutions of ribonuclease. Biophys Chem 108:89-100.
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