PharmPK Discussion - Plasma protein binding utilizing 96-wells micro-dialysis plates

• On 28 Jun 2007 at 15:45:14, "Michelle Rudek" (mrudek2.-at-.jhmi.edu) sent the message
  

  
  The following message was posted to: PharmPK
  
  I have a compound that we are planning on doing in vitro PPB
  experiments on.  The compound is proposed to be 'sticky.'  I have
  contacted Harvard Apparatus with their recommendation to pre-treat
  with a surfactant.  They had recommended either Triton X or PEG with
  a preference for Triton X (up to a final concentration of 0.1%, v/
  v).  Has anyone utilized PEG in PPB experiments to pre-treat for non-
  specific?  If so, what concentration (%, v/v) did you utilize?
  
  Thanks,
  Michelle
  
  

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• On 29 Jun 2007 at 09:35:25, "Sheldon, Adrian" (Adrian.Sheldon.aaa.crl.com) sent the message
  

  
  The following message was posted to: PharmPK
  
  One approach to consider is using a different device.  My lab had a poor
  experience with the Harvard plate (including leaking wells), and use a
  HTDialysis Teflon device with good results (Teflon may reduce NSB of
  your sticky compound).  Note that if it is hydrophobic/highly bound, you
  may need a long time to reach equilibrium.  The RED device is also
  well-regarded.
  Another approach we have successfully used is ultracentrifugation.
  Again, you'd need to perform controls to assess recovery.
  Adrian Sheldon
  Charles River Labs, in Vitro ADMET
  
  

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• On 30 Jun 2007 at 01:15:34, stanleycotl.-at-.aol.com sent the message
  

  
  I used the Harvard plates. Newer one's seemed to solve the problem of
  leaking. What I did was to calculate the mass balance. At equilibrium
  amt in buffer + amt in plasma divided by initial concentration X 100.
  Yes, it is important to run a timed study to determine time of
  equilibrium. Stanley Cotler
  
  

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