<html><head><base href="x-msg://384/"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; "><font class="Apple-style-span" color="#950f0b">Hi John</font><div><font class="Apple-style-span" color="#950f0b"><br></font></div><div><font class="Apple-style-span" color="#950f0b">I have done SV on many IgG samples. 1.5 if a fair average for f/f0 - but IgG's are (mostly) flexible structures, their level of glycosylation is indeed variable, so for choice - if the exact value matters to you - it's best to measure it.</font></div><div><font class="Apple-style-span" color="#950f0b"><br></font></div><div><font class="Apple-style-span" color="#950f0b">Kind regards</font></div><div><font class="Apple-style-span" color="#950f0b"><br></font></div><div><font class="Apple-style-span" color="#950f0b">Arthur</font></div><div><font class="Apple-style-span" color="#950f0b"><br></font></div><div><font class="Apple-style-span" color="#950f0b">Arthur J Rowe<br>125 Uplands Road<br>Oadby<br>Leicester<br>LE2 4NW<br><br>Tel: +44 116 2714502<br><a href="mailto:arthur.rowe@connectfree.co.uk">arthur.rowe@connectfree.co.uk</a></font></div><div><br><div><div>On 14 Jan 2014, at 17:38, John Sumida wrote:</div><br class="Apple-interchange-newline"><blockquote type="cite"><span class="Apple-style-span" style="border-collapse: separate; font-family: Helvetica; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: 2; text-align: -webkit-auto; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-border-horizontal-spacing: 0px; -webkit-border-vertical-spacing: 0px; -webkit-text-decorations-in-effect: none; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px; font-size: medium; "><div lang="EN-US" link="blue" vlink="purple"><div class="WordSection1" style="page: WordSection1; "><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; ">Natalia and Mattia,<o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; "><o:p> </o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; ">Thank you for your responses. I gather that in general these values are not measured for the CFCA analysis and that perhaps there is not sufficient reason to do so given the precision of the application? I wonder how variably glycosylated proteins would affect the accuracy of this technique?<o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; "><o:p> </o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; ">Thank you both again for taking the time to respond.<o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; "><o:p> </o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; "><o:p> </o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; ">Best regards,<o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; ">John Sumida, Ph.D.<o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: rgb(31, 73, 125); "><a href="http://depts.washington.edu/cidb4bio/index.shtml" style="color: blue; text-decoration: underline; ">Analytical Biopharmacy Core Facility</a><o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; ">University of Washington<o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: rgb(31, 73, 125); "><a href="http://www.moles.washington.edu/" style="color: blue; text-decoration: underline; ">Molecular Engineering & Sciences Institute, G22</a><o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; ">3946 West Stevens Way NE<o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; ">Seattle WA 98195-1651<o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; "><o:p> </o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; "><o:p> </o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><span style="font-size: 11pt; font-family: 'Courier New'; color: blue; "><o:p> </o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><b><span style="font-size: 10pt; font-family: Tahoma, sans-serif; ">From:</span></b><span style="font-size: 10pt; font-family: Tahoma, sans-serif; "><span class="Apple-converted-space"> </span>Mattia Rocco [mailto:mattia.rocco@hsanmartino.it]<span class="Apple-converted-space"> </span><br><b>Sent:</b><span class="Apple-converted-space"> </span>Tuesday, January 14, 2014 1:17 AM<br><b>To:</b><span class="Apple-converted-space"> </span>Markova, Natalia (GE Healthcare); John Sumida;<span class="Apple-converted-space"> </span><a href="mailto:rasmb@rasmb.org" style="color: blue; text-decoration: underline; ">rasmb@rasmb.org</a><br><b>Subject:</b><span class="Apple-converted-space"> </span>Re: [RASMB] SPR query about CFCA applications<o:p></o:p></span></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><o:p> </o:p></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; "><br>The f/f0 Natalia mentions is for almost spherical shape.... if you are dealing with an antibody, then the 1.5 value might be more appropriate (but I haven't done any calculations on it... ;-) )<br><br>Best - Mattia<br><br>At 08:13 AM 1/14/2014 +0000, Markova, Natalia (GE Healthcare) wrote:<br><br><o:p></o:p></div><div style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; margin-top: 0in; margin-bottom: 0.0001pt; ">Content-Language: en-US<br>Content-Type: multipart/alternative;<br> boundary="_000_41C5780FD432C74DB68C588BE66E34F614EF6B76BUDURBPA08e2kad_"<br><br>Hi John,<br><br> <br><br>We use as default f/f0 factor for globular proteins, which is 1.2 (not 1.5). However, user can freely enter any value for this factor in Web diffusion coefficient calculator (<a href="https://www.biacore.com/lifesciences/Application_Support/online_support/Diffusion_Coefficient_Calculator/index.html?section=lifesciences&realsection=lifesciences" style="color: blue; text-decoration: underline; ">https://www.biacore.com/lifesciences/Application_Support/online_support/Diffusion_Coefficient_Calculator/index.html?section=lifesciences&realsection=lifesciences</a>)<br><br>You need to be registered on the Biacore website to access it.<br><br> <br><br>Best regards,<br><br> <br><br>Natalia<br><br> <br><br><b><i>Natalia Markova (PhD)<span class="Apple-converted-space"> </span><br></i></b><br><i>GE Healthcare<br></i><br><i>R&D Life Sciences<br></i><br><i>Rapsgatan 23<br></i><br><i>751 84 Uppsala<br></i><br><i>Sweden<br></i><br><i> <br></i><br>M +46 727 33 58 80<br><br>E <span class="Apple-converted-space"> </span><a href="mailto:natalia.markova@ge.com" style="color: blue; text-decoration: underline; ">natalia.markova@ge.com</a><br><br><i> <br></i><br><a href="http://www.biacore.com/" style="color: blue; text-decoration: underline; ">www.biacore.com</a><br><a href="https://exchange1.ad.slu.se/owa/redir.aspx?C=61606803db87492ba25280057e5d208e&URL=http%3a%2f%2fwww.microcal.com%2f" style="color: blue; text-decoration: underline; ">www.microcal.com</a><br><br><br><br>Disclaimer<br><br>The information contained in this e-mail is intended solely for the use of the addressee and contains information that is confidential and may be legally privileged. Access to this e-mail by anyone else is unauthorised. If you are not the intended recipient, your use, retention, copy, any disclosure, dissemination or any action taken or omitted to be taken in reliance of this communication is strictly prohibited and may be unlawful. If you have received this communication in error, please notify us immediately by phone. Thank you!<br><br> <br><br> <br><br> <br><br> <br><br><b>From:</b><span class="Apple-converted-space"> </span><a href="mailto:rasmb-bounces@list.rasmb.org" style="color: blue; text-decoration: underline; ">rasmb-bounces@list.rasmb.org</a><span class="Apple-converted-space"> </span>[<a href="mailto:rasmb-bounces@list.rasmb.org" style="color: blue; text-decoration: underline; ">mailto:rasmb-bounces@list.rasmb.org</a>]<span class="Apple-converted-space"> </span><b>On Behalf Of<span class="Apple-converted-space"> </span></b>John Sumida<br><b>Sent:</b><span class="Apple-converted-space"> </span>den 14 januari 2014 00:20<br><b>To:</b><span class="Apple-converted-space"> </span><a href="mailto:rasmb@rasmb.org" style="color: blue; text-decoration: underline; ">rasmb@rasmb.org</a><br><b>Subject:</b><span class="Apple-converted-space"> </span>[RASMB] SPR query about CFCA applications<br><br> <br><br>Dear RASMB,<br><br> <br><br>This question is in regards to an SPR application. We are currently investigating the use calibration free concentration analysis (CFCA) to monitor the stability of macromolecular preps over time in terms of their activity. Input parameters for this analysis are ff0, viscosity, MW and D and we are using AUC to better determine both. Current applications seem to assume ff0=1.5, which may not be correct for monoclonal preps based on my reading of the literature.<br><br> <br><br>My query.<br><br>1. Is it currently the common practice to measure these quantities and if so what is the preferred method to do so?<br><br> <br><br>Thank you all and Happy New Year,<br><br> <br><br>John Sumida, Ph.D.<br><br><a href="http://depts.washington.edu/cidb4bio/index.shtml" style="color: blue; text-decoration: underline; ">Analytical Biopharmacy Core Facility</a><br><br>University of Washington<br><br><a href="http://www.moles.washington.edu/" style="color: blue; text-decoration: underline; ">Molecular Engineering & Sciences Institute, G22</a><br><br>3946 West Stevens Way NE<br><br>Seattle WA 98195-1653<br><br> <br>_______________________________________________<br>RASMB mailing list<br><a href="mailto:RASMB@list.rasmb.org" style="color: blue; text-decoration: underline; ">RASMB@list.rasmb.org</a><br><a href="http://list.rasmb.org/listinfo.cgi/rasmb-rasmb.org" style="color: blue; text-decoration: underline; ">http://list.rasmb.org/listinfo.cgi/rasmb-rasmb.org</a><o:p></o:p></div><p style="margin-right: 0in; margin-left: 0in; font-size: 12pt; font-family: 'Times New Roman', serif; "><span style="font-size: 10pt; font-family: Arial, sans-serif; "><br></span><span style="font-size: 10pt; font-family: Verdana, sans-serif; ">ATTENZIONE.<br>Il presente messaggio ed i suoi allegati devono intendersi ad uso esclusivo dei suoi destinatari e sono confidenziali. 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Mail is not a secure protocol, therefore IST will not be liable<br>for interception or amendment of this message.</i></span><o:p></o:p></p></div>_______________________________________________<br>RASMB mailing list<br><a href="mailto:RASMB@list.rasmb.org" style="color: blue; text-decoration: underline; ">RASMB@list.rasmb.org</a><br><a href="http://list.rasmb.org/listinfo.cgi/rasmb-rasmb.org" style="color: blue; text-decoration: underline; ">http://list.rasmb.org/listinfo.cgi/rasmb-rasmb.org</a><br></div></span></blockquote></div><br><div>
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