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</o:shapelayout></xml><![endif]--></head><body lang=EN-US link=blue vlink=purple><div class=WordSection1><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Hello all,<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>The ViscoStar could very well be employed to measure absolute viscosity directly, with minor changes to the software. The question to the group is, what range and accuracy are required for the studies you envision? I’ve asked Richard off-line but might as well get a consensus, then let you know if it is feasible. <o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Alternatively, for protein characterization under moderate to low concentrations, you could use the specific viscosity reported now in ASTRA, multiply by concentration and add it to the solvent viscosity for an absolute value. If there is sufficient interest we could modify the software quite easily to do that for you.<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Thoughts?<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Best,<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Dan<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><div><div style='border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in'><p class=MsoNormal><b><span style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>From:</span></b><span style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> rasmb-bounces@list.rasmb.org [mailto:rasmb-bounces@list.rasmb.org] <b>On Behalf Of </b>Richard Kingston<br><b>Sent:</b> Thursday, June 06, 2013 8:53 PM<br><b>To:</b> David Scott<br><b>Cc:</b> rasmb@rasmb.org<br><b>Subject:</b> Re: [RASMB] Viscometer<o:p></o:p></span></p></div></div><p class=MsoNormal><o:p> </o:p></p><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><p class=MsoNormal>Thanks for the many informative replies, both on- and off- list.<o:p></o:p></p><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal>The consensus seems to be that differential (pressure-imbalance) viscometers like the Viscostar are well suited to the measurement of protein intrinsic viscosities, and there are several in commercial production. However these instruments, by design, measure only relative viscosities <o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal>Reframing my original question, I was really wondering if it were possible to find an instrument which could measure absolute solution viscosities with sufficient accuracy to allow protein characterisation (and could therefore do double duty, being able to measure solution viscosity for other biophysical applications). However for the reasons outlined by Arthur, and as attested below by Dave, it might be difficult to achieve good results this way.<o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal>Best,<o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal>Richard<o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><div><p class=MsoNormal>On 7/06/2013, at 5:56 AM, David Scott <<a href="mailto:David.Scott@nottingham.ac.uk">David.Scott@nottingham.ac.uk</a>> wrote:<o:p></o:p></p></div><p class=MsoNormal><br><br><o:p></o:p></p><div><p class=MsoNormal>Hello All in RASMB land,<o:p></o:p></p><div><p class=MsoNormal><o:p> </o:p></p><div><p class=MsoNormal>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:<o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal><span class=apple-style-span><span style='font-family:"Arial","sans-serif"'>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. </span></span><span class=apple-style-span><i><span style='font-size:13.5pt;font-family:"Arial","sans-serif"'>J. Biol. Chem</span></i></span><span class=apple-style-span><span style='font-size:13.5pt;font-family:"Arial","sans-serif"'>. </span></span><span class=slug-vol><b><span lang=EN style='font-size:13.5pt;font-family:"Arial","sans-serif"'>285</span></b></span><span class=slug-vol><span lang=EN style='font-size:13.5pt;font-family:"Arial","sans-serif"'> </span></span><span class=slug-pages3><span lang=EN style='font-size:13.5pt;font-family:"Arial","sans-serif"'>15440-15449</span></span><o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><div><div><p class=MsoNormal>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. <o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal>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.<o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal>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...).<o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal>Best<o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal>Dave Scott.<o:p></o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal><o:p> </o:p></p></div><div><p class=MsoNormal>On 6 Jun 2013, at 17:22, Razinkov, Vladimir wrote:<o:p></o:p></p></div><p class=MsoNormal><br><br><o:p></o:p></p><div><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Dear all,</span><o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'> </span><o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>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.</span><o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'> </span><o:p></o:p></p></div><div><p class=MsoNormal style='line-height:17.4pt;background:white'><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'><a href="http://www.ncbi.nlm.nih.gov/pubmed/19995543" title="Analytical biochemistry."><span style='color:#660066'>Anal Biochem.</span></a> 2010 Apr 1;399(1):141-3. doi: 10.1016/j.ab.2009.12.003. Epub 2009 Dec 6.</span><o:p></o:p></p></div><p class=MsoNormal style='margin-bottom:4.5pt;background:white;background-image:initial;background-attachment:initial;background-origin: initial;background-clip: initial;background-position:initial initial;background-repeat:initial initial'><b><span style='font-size:15.0pt;font-family:"Arial","sans-serif"'>High-throughput dynamic light scattering method for measuring viscosity of concentrated protein solutions.</span></b><o:p></o:p></p><div><p class=MsoNormal style='background:white'><span style='font-size:11.0pt;font-family:"Arial","sans-serif"'><a href="http://www.ncbi.nlm.nih.gov/pubmed?term=He%20F%5BAuthor%5D&cauthor=true&cauthor_uid=19995543"><span style='color:#660066'>He F</span></a>, <a href="http://www.ncbi.nlm.nih.gov/pubmed?term=Becker%20GW%5BAuthor%5D&cauthor=true&cauthor_uid=19995543"><span style='color:#660066'>Becker GW</span></a>, <a href="http://www.ncbi.nlm.nih.gov/pubmed?term=Litowski%20JR%5BAuthor%5D&cauthor=true&cauthor_uid=19995543"><span style='color:#660066'>Litowski JR</span></a>, <a href="http://www.ncbi.nlm.nih.gov/pubmed?term=Narhi%20LO%5BAuthor%5D&cauthor=true&cauthor_uid=19995543"><span style='color:#660066'>Narhi LO</span></a>, <a href="http://www.ncbi.nlm.nih.gov/pubmed?term=Brems%20DN%5BAuthor%5D&cauthor=true&cauthor_uid=19995543"><span style='color:#660066'>Brems DN</span></a>, <a href="http://www.ncbi.nlm.nih.gov/pubmed?term=Razinkov%20VI%5BAuthor%5D&cauthor=true&cauthor_uid=19995543"><span style='color:#660066'>Razinkov VI</span></a>.</span><o:p></o:p></p></div><p class=MsoNormal style='margin-bottom:7.7pt;background:white;background-image:initial;background-attachment:initial;background-origin: initial;background-clip: initial;background-position:initial initial;background-repeat:initial initial'><b><span style='font-size:11.0pt;font-family:"Arial","sans-serif";color:#724128'>Source</span></b><o:p></o:p></p><p class=MsoNormal style='margin-bottom:6.0pt;line-height:13.1pt;background:white;background-image:initial;background-attachment:initial;background-origin: initial;background-clip: initial;background-position:initial initial;background-repeat:initial initial'><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'>Formulation and Analytical Resources, Amgen Inc., Seattle, WA 98119, USA.</span><o:p></o:p></p><div><p class=MsoNormal style='background:white'><b><span style='font-size:11.0pt;font-family:"Arial","sans-serif";color:#985735'>Abstract</span></b><o:p></o:p></p></div><p class=MsoNormal style='margin-bottom:6.0pt;line-height:12.75pt;background:white;background-image:initial;background-attachment:initial;background-origin: initial;background-clip: initial;background-position:initial initial;background-repeat:initial initial'><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'>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</span><o:p></o:p></p><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'> </span><o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'> </span><o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Regards</span><o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'> </span><o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Vladimir Razinkov</span><o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'> </span><o:p></o:p></p></div><div><div style='border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in;border-width:initial;border-color:initial'><div><p class=MsoNormal><b><span style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>From:</span></b><span class=apple-converted-space><span style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> </span></span><span style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'><a href="mailto:rasmb-bounces@list.rasmb.org">rasmb-bounces@list.rasmb.org</a><span class=apple-converted-space> </span>[mailto:rasmb-<a href="mailto:bounces@list.rasmb.org">bounces@list.rasmb.org</a>]<span class=apple-converted-space> </span><b>On Behalf Of<span class=apple-converted-space> </span></b>Richard Kingston<br><b>Sent:</b><span class=apple-converted-space> </span>Wednesday, June 05, 2013 9:15 PM<br><b>To:</b><span class=apple-converted-space> </span><a href="mailto:rasmb@rasmb.org">rasmb@rasmb.org</a><br><b>Subject:</b><span class=apple-converted-space> </span>[RASMB] Viscometer</span><o:p></o:p></p></div></div></div><div><p class=MsoNormal> <o:p></o:p></p></div><div><div><p class=MsoNormal> <o:p></o:p></p></div></div><div><div><p class=MsoNormal>Dear all,<o:p></o:p></p></div></div><div><div><p class=MsoNormal> <o:p></o:p></p></div></div><div><div><p class=MsoNormal>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. <o:p></o:p></p></div></div><div><div><p class=MsoNormal> <o:p></o:p></p></div></div><div><div><p class=MsoNormal>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.<o:p></o:p></p></div></div><div><div><p class=MsoNormal> <o:p></o:p></p></div></div><div><div><p class=MsoNormal>Many thanks,<o:p></o:p></p></div></div><div><div><p class=MsoNormal> <o:p></o:p></p></div></div><div><div><p class=MsoNormal>Richard<o:p></o:p></p></div></div><div><div><p class=MsoNormal> <o:p></o:p></p></div></div><div><div><div><p class=MsoNormal><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif"'>Richard Kingston, PhD.</span><o:p></o:p></p></div></div><div><div><p class=MsoNormal><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif"'>School of Biological Sciences</span><o:p></o:p></p></div></div><div><div><p class=MsoNormal><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif"'>The University of Auckland</span><o:p></o:p></p></div></div><div><div><p class=MsoNormal><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif"'>New Zealand</span><o:p></o:p></p></div></div><div><div><p class=MsoNormal><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif"'> </span><o:p></o:p></p></div></div><div><div><p class=MsoNormal><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif"'>website: <a href="http://persephone.sbs.auckland.ac.nz/richard/lab/">http://persephone.sbs.auckland.ac.nz/richard/lab/</a></span><o:p></o:p></p></div></div><div><div><p class=MsoNormal><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif"'> </span><o:p></o:p></p></div></div><p class=MsoNormal style='margin-bottom:12.0pt'><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif"'> </span><o:p></o:p></p></div><div><p class=MsoNormal> <o:p></o:p></p></div><p class=MsoNormal>_______________________________________________<br>RASMB mailing list<br><a href="mailto:RASMB@list.rasmb.org">RASMB@list.rasmb.org</a><br><a href="http://list.rasmb.org/listinfo.cgi/rasmb-rasmb.org">http://list.rasmb.org/listinfo.cgi/rasmb-rasmb.org</a><o:p></o:p></p></div></div><p class=MsoNormal><o:p> </o:p></p></div></div><p class=MsoNormal><o:p> </o:p></p><p>This message and any attachment are intended solely for the addressee and may contain confidential information. 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