[RASMB] SPR query about CFCA applications

Tue Jan 14 09:38:48 PST 2014

Natalia and Mattia,

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?

Thank you both again for taking the time to respond.

Best regards,

John Sumida, Ph.D.

Analytical Biopharmacy Core Facility
<http://depts.washington.edu/cidb4bio/index.shtml> 

University of Washington

Molecular Engineering  <http://www.moles.washington.edu/> & Sciences
Institute, G22

3946 West Stevens Way NE

Seattle WA 98195-1651

From: Mattia Rocco [mailto:mattia.rocco at hsanmartino.it] 
Sent: Tuesday, January 14, 2014 1:17 AM
To: Markova, Natalia (GE Healthcare); John Sumida; rasmb at rasmb.org
Subject: Re: [RASMB] SPR query about CFCA applications

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... ;-) )

Best - Mattia

At 08:13 AM 1/14/2014 +0000, Markova, Natalia (GE Healthcare) wrote:

Content-Language: en-US
Content-Type: multipart/alternative;

boundary="_000_41C5780FD432C74DB68C588BE66E34F614EF6B76BUDURBPA08e2kad_"

Hi John,

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
(https://www.biacore.com/lifesciences/Application_Support/online_support/Dif
fusion_Coefficient_Calculator/index.html?section=lifesciences
<https://www.biacore.com/lifesciences/Application_Support/online_support/Dif
fusion_Coefficient_Calculator/index.html?section=lifesciences&realsection=li
fesciences> &realsection=lifesciences)

You need to be registered on the Biacore website to access it.

Best regards,

Natalia

Natalia Markova (PhD) 

GE Healthcare

R&D Life Sciences

Rapsgatan 23

751 84 Uppsala

Sweden

M  +46 727 33 58 80

E   natalia.markova at ge.com

www.biacore.com <http://www.biacore.com/> 
www.microcal.com
<https://exchange1.ad.slu.se/owa/redir.aspx?C=61606803db87492ba25280057e5d20
8e&URL=http%3a%2f%2fwww.microcal.com%2f> 

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From: rasmb-bounces at list.rasmb.org [mailto:rasmb-bounces at list.rasmb.org] On
Behalf Of John Sumida
Sent: den 14 januari 2014 00:20
To: rasmb at rasmb.org
Subject: [RASMB] SPR query about CFCA applications

Dear RASMB,

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.

My query.

1.       Is it currently the common practice to measure these quantities and
if so what is the preferred method to do so?

Thank you all and Happy New Year,

John Sumida, Ph.D.

Analytical Biopharmacy Core Facility
<http://depts.washington.edu/cidb4bio/index.shtml> 

University of Washington

Molecular Engineering  <http://www.moles.washington.edu/> & Sciences
Institute, G22

3946 West Stevens Way NE

Seattle WA 98195-1653

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