[RASMB] Viscometer

[David Scott]

Hello All in RASMB land,

We have both the viscostar and the Anton Paar viscometer in the NCMH, and have used both to try and attain viscosity increments. In practice, only the viscostar actually works at reasonable concentrations (c.a. 1 mg/ml) for proteins. We've used this to look at conformations of intrinsically disordered proteins:

Rajeskar, K.V., Muntaha, S.T., Tame, J.R.H., Wharton, C.M., Thomas, C.M., White, S.A., Hyde, E.I. and Scott, D.J. (2010). Order and disorder in the domain organisation of the plasmid partition protein KorB. J. Biol. Chem. 285 15440-15449

It gives very nice results, especially coupled to SEC-MALLS. I would also add to Mattia's point that using absorbance to measure your protein concentration is so much more accurate than refractive index increments.

The Anton Paar system gives very good (3 d.p.) precision measurements of solution viscosities which are essential for accurate corrections of sedimentation velocity data, but it's use beyond this is limited, in my experience. I know others in the NCMH have tried to push it further with carbohydrate systems, but they tend to be more abundant in yields than proteins.

The only downside to the viscostar is a tendency to blockages, so needs to be used on a fairly constant basis, and washed through regularly. However, despite that caveat, I have been impressed with it (when it is working properly...).

Best

Dave Scott.







On 6 Jun 2013, at 17:22, Razinkov, Vladimir wrote:

Dear all,

Another rather simple and high throughput  method to measure viscosity of proteins at low volumes (~ 35 uL with 384-well plate) using DLS plate reader.

Anal Biochem.<http://www.ncbi.nlm.nih.gov/pubmed/19995543> 2010 Apr 1;399(1):141-3. doi: 10.1016/j.ab.2009.12.003. Epub 2009 Dec 6.
High-throughput dynamic light scattering method for measuring viscosity of concentrated protein solutions.
He F<http://www.ncbi.nlm.nih.gov/pubmed?term=He%20F%5BAuthor%5D&cauthor=true&cauthor_uid=19995543>, Becker GW<http://www.ncbi.nlm.nih.gov/pubmed?term=Becker%20GW%5BAuthor%5D&cauthor=true&cauthor_uid=19995543>, Litowski JR<http://www.ncbi.nlm.nih.gov/pubmed?term=Litowski%20JR%5BAuthor%5D&cauthor=true&cauthor_uid=19995543>, Narhi LO<http://www.ncbi.nlm.nih.gov/pubmed?term=Narhi%20LO%5BAuthor%5D&cauthor=true&cauthor_uid=19995543>, Brems DN<http://www.ncbi.nlm.nih.gov/pubmed?term=Brems%20DN%5BAuthor%5D&cauthor=true&cauthor_uid=19995543>, Razinkov VI<http://www.ncbi.nlm.nih.gov/pubmed?term=Razinkov%20VI%5BAuthor%5D&cauthor=true&cauthor_uid=19995543>.
Source
Formulation and Analytical Resources, Amgen Inc., Seattle, WA 98119, USA.
Abstract
We propose a new method to measure the viscosity of concentrated protein solutions in a high-throughput format. This method measures the apparent hydrodynamic radius of polystyrene beads with known sizes using a dynamic light scattering (DLS) system with a microplate reader. Glycerol solution viscosities obtained by the DLS method were in good agreement with those reported in the literature. Viscosity of the solutions of two monoclonal antibody molecules was acquired using both DLS and cone-and-plate techniques, and the results were comparable. The DLS method described here has the potential to be used in many aspects of protein characterization


Regards

Vladimir Razinkov

From: rasmb-bounces at list.rasmb.org<mailto:rasmb-bounces at list.rasmb.org> [mailto:rasmb-bounces at list.rasmb.org] On Behalf Of Richard Kingston
Sent: Wednesday, June 05, 2013 9:15 PM
To: rasmb at rasmb.org<mailto:rasmb at rasmb.org>
Subject: [RASMB] Viscometer


Dear all,

I've become interested in measuring the viscosity of protein solutions at varying temperature,  and wondered if anyone on the list could advise on the best way to do this using commercially available instrumentation. While there are many  ways to measure viscosity, for protein applications minimizing the required sample volume is pretty critical. I note that  Anton Paar sell an instrument that measures both dynamic viscosity and density  (SVM 3000 Stabinger viscometer). If you skip the density measurement, the sample requirements look almost "manageable". Cambridge also sell a micro-sample viscometer (Viscolab 5000) which uses microlitre volumes.

I'm having difficulty tracking down people that might have used these or similar instruments, and could comment on the practicalities for protein work. If anyone can offer advice, either on- or off-list, it would be appreciated.

Many thanks,

Richard

Richard Kingston, PhD.
School of Biological Sciences
The University of Auckland
New Zealand

website: http://persephone.sbs.auckland.ac.nz/richard/lab/



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