[RASMB] AUC of protein formulation

[Arthur Rowe]

Hi John

This is an area where I do quite a bit of work, as you may guess. My first
thought is "why do you need to know the solvent parameters, given that what
you are looking for is simply degree of oligomerisation?". The lack of such
knowledge may indeed slightly mess up the absolute values in a c(M) profile,
but given that you almost certainly know the formula weight of your
monomeric product, who cares? What you get will surely be good enough to say
"that peak is the monomer!"

However, a second thought comes to mind, again based upon experience, when I
see that you have "surface-active excipients" present. These may well form
micellar structures in solution, into which the proteins monomers etc are
incorporated. But then again, the amount of such excipients added may be
minimal, and below the CMC (critical micelle concentration). There are ways
of handling such systems, and I have recently published some novel
approaches which might be applicable in your case. I will happily enlarge on
this topic if it seems relevant to your system.

Kind regards

Arthur

--
*******************************************************
Arthur J Rowe
Professor of Biomolecular Technology
NCMH Business Centre
University of Nottingham
School of Biosciences
Sutton Bonington
Leicestershire LE12 5RD   UK

Tel:        +44 (0)115 951 6156
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Fax:        +44 (0)115 951 6157
email:      arthur.rowe at nottingham.ac.uk
Web:        www.nottingham.ac.uk/ncmh/business
*******************************************************


From: john.b.babetas at gskbio.com
Date: Mon, 3 Nov 2008 09:12:33 -0500
To: rasmb at rasmb.bbri.org
Subject: [RASMB] AUC of protein formulation



To whom it may concern,

I am in the process of using Sedimentation Velocity- Analytical
Ultracentrifugation to evaluate the aggregation of a protein formulation.
The formulation is complex containing Sugars, Monothioglycerol and
surfactant excipients. I wanted to know what needs to be done in terms of
Data processing to deal with a complex matrix as this one. When i use
sednterp not all the components of the solvent are there. So i cannot
determine the density and viscosity with exactitude. I know that the
inhomogeneous solvent option deals with dynamic gradients coming from one
co-solute and not many. I wanted to know what are my options to overcome
this problem. 

John
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