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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:black">For viscosity measurements of small volume protein solutions we use Wyatt DynaPro plate reader and 384-well plate. It takes ~ 35 uL of protein sample. Diffusion
coefficient of added into solution beads with known radius is measured and converted to viscosity. See our original article:
</span><b><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:black;background:white">He, Feng, et al. "High-throughput dynamic light scattering method for measuring viscosity of concentrated protein solutions."<span class="apple-converted-space"> </span><i>Analytical
biochemistry</i>399.1 (2010): 141-143</span></b><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:black;background:white">. It is good tool for quick selection of viscous candidates.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:black;background:white">Protein-bead interactions and subsequent aggregation can be a problem. But, based on our experience, it is rare for antibodies.
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:black;background:white"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:black;background:white">Vladimir Razinkov<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:#222222;background:white"><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>
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<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 [mailto:rasmb-bounces@list.rasmb.org]
<b>On Behalf Of </b>Stephen Harding<br>
<b>Sent:</b> Wednesday, June 03, 2015 1:45 PM<br>
<b>To:</b> John Sumida; 'David Hayes'; 'RASMB'<br>
<b>Subject:</b> Re: [RASMB] Viscometer technology<o:p></o:p></span></p>
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<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:11.0pt;color:black">John: interest in protein intrinsic viscosity is that it is a far more sensitive function of conformation in solution compared with the sedimentation coefficient. In addition, interest
in absolute viscosity is important in for example monoclonal antibody based formulations – high viscosity is a problem in terms of administration.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:11.0pt;color:black">For conformation work, problem in the past has been that of sample heterogeneity – sedimentation velocity could resolve away aggregates etc, viscometry couldn’t – not until the advent
of the differential pressure viscometers (measuring relative viscosity by pressure tranducers rather than photosensors) – which could be linked onto SEC columns and concentration detectors. These instruments (Wyatt Viscostar or the Malvern Viscotek) are so
sensitive that they can measure relative viscosity at concentrations low enough so that a concentration extrapolation of the reduced viscosity to zero concentration c=0 are not necessary (there are other tricks as well!).<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:11.0pt;color:black">There have been difficulties largely arising from corrosion of the transducers after being exposed to salt ions in aqueous solution but they seem to have got round that now by appropriate
coating - indeed its nice instrumentation.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:11.0pt;color:black">David – yes the rolling ball viscometer (Anton Paar) although its been good for looking at polysaccharide solutions – the ability to vary the angle of flow changes the shear rate so
you can monitor for non-Newtonian behaviour as with Cone and Plate (but much lower volumes) is really useful….. but bad for protein solution work. That was the view 5 years ago at the great BITC meeting in Galveston – and is still the same. The Anton-Paar
team were really helpful when we approached them about silanization and this improved things but not really enough. For polysaccharides yes, proteins no.
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:11.0pt;color:black">For intrinsic viscosity measurement on proteins we would probably still go for the standard Ostwald U-tube capillary (1st choice) or if polydispersity is suspected the Viscostar (2<sup>nd</sup>
choice). For polysaccharide/ glycoconjugate work, the Viscostar would be 1<sup>st</sup> choice.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:11.0pt;color:black">The new Malvern Viscosizer is based on a different principle altogether. Malvern for some time have been trying to see if dynamic light scattering on particles of known size can be
used as a measure of the viscosity of a medium from the relation of the diffusion coefficient with viscosity, and the Taylor dispersion based instrument I guess is a development from that so it would be interesting to see how it goes. The problems of protein
adhesion sounds ominously familiar! It looks to me anyway as if its intended for absolute viscosity measurement rather than being accurate enough for intrinsic viscosity.
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:11.0pt;color:black">All best,
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:11.0pt;color:black">Steve<o:p></o:p></span></p>
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<p class="MsoNormal"><i><span lang="DE-CH" style="font-family:"Verdana","sans-serif";color:red">________________________</span></i><span lang="DE-CH" style="font-family:"Verdana","sans-serif";color:red"><o:p></o:p></span></p>
<p class="MsoNormal"><span lang="DE-CH" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black">Steve Harding DSc (Oxon), RFOO (Norway)<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="DE-CH" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black">Professor of Applied Biochemistry,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="DE-CH" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black">University of Nottingham,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="DE-CH" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black">School of Biosciences,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="DE-CH" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black">Sutton Bonington LE12 5RD, England<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="DE-CH" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black">(44) (0) 78110 90635<o:p></o:p></span></p>
<p class="MsoNormal" style="margin-bottom:12.0pt"><u><span lang="DE-CH" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black"><a href="http://www.nottingham.ac.uk/Biosciences/People/steve.harding"><span style="color:black">www.nottingham.ac.uk/Biosciences/People/steve.harding</span></a><o:p></o:p></span></u></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black">National Centre for Macromolecular Hydrodynamics (NCMH):<b><o:p></o:p></b></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black"><a href="http://www.nottingham.ac.uk/ncmh"><span style="color:black">www.nottingham.ac.uk/ncmh</span></a><o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black">Genetic Survey of North West England:<b><br>
</b><a href="http://www.nottingham.ac.uk/-sczsteve/survey.htm"><span style="color:black">www.nottingham.ac.uk/-sczsteve/survey.htm</span></a><o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:black">(BBSRC Watson-Crick Anniversary Award)<o:p></o:p></span></p>
<p class="MsoNormal" style="margin-bottom:12.0pt"><i><span lang="DE-CH" style="font-family:"Verdana","sans-serif";color:yellow">________________________</span></i><span lang="DE-CH" style="font-size:6.0pt;font-family:"Verdana","sans-serif";color:yellow"><o:p></o:p></span></p>
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<p class="MsoNormal"><span lang="EN-GB" style="font-size:11.0pt;color:black"><o:p> </o:p></span></p>
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<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 [<a href="mailto:rasmb-bounces@list.rasmb.org">mailto:rasmb-bounces@list.rasmb.org</a>]
<b>On Behalf Of </b>John Sumida<br>
<b>Sent:</b> 03 June 2015 17:32<br>
<b>To:</b> 'David Hayes'; 'RASMB'<br>
<b>Subject:</b> Re: [RASMB] Viscometer technology<o:p></o:p></span></p>
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<p class="MsoNormal"><span lang="EN-GB"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue">Dear David,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue">Your post piqued my curiosity. Please forgive my ignorance on the topic, but why is protein viscosity something one would be interested in? And for those who make these
types of measurements, what is the variability in protein viscosity?<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue">Thank you all for your comments and responses, now and in the past. This bulletin board has been a great learning tool.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue">Best regards,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue">John Sumida<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue">University of Washington<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Courier New";color:blue"><o:p> </o:p></span></p>
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<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 [<a href="mailto:rasmb-bounces@list.rasmb.org">mailto:rasmb-bounces@list.rasmb.org</a>]
<b>On Behalf Of </b>David Hayes<br>
<b>Sent:</b> Wednesday, June 03, 2015 5:23 AM<br>
<b>To:</b> RASMB<br>
<b>Subject:</b> [RASMB] Viscometer technology<o:p></o:p></span></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black">Hi all (especially hoping Steve Harding is active),<o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black"> <o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black">I recently saw the Malvern viscosizer. It is a capillary viscometer and taylor dispersion instrument. (Taylor dispersion looks at the diffusion pattern
of a small plug of sample in a capillary). Because of the small volumes used, it could be useful for screening early limited supply proteins for viscosity problems.<o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black"> <o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black">Are there any general thoughts on the technology?<o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black"> <o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black">I remember Steve Harding telling us at BITC that you never use rolling ball viscometers on proteins: the proteins stick to the ball.<o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black">Recently we found out here that the regular Anton Paar cone and plate rheometers sometimes need PS-80 in protein samples or the air water interface
around the outside edge can predominate the viscosity measurements.<o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black"> <o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black">I did ask whether most samples just stick to the capillary and disappear or run anomalously, but the answer was they had a super coating so most things
won’t stick. This coating makes the capillary impossible to clean (so you buy more capillaries from Malvern). And that the taylor dispersion fitting will tell you if and approximately how much the sample interacts with the capillary walls.<o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black"> <o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black">Kind Regards,<o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black"> <o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black">David Hayes<o:p></o:p></span></p>
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<p class="MsoNormal" style="background:white"><span style="font-family:"Helvetica","sans-serif";color:black"><o:p> </o:p></span></p>
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