PharmPK Discussion - Non specific binding equation

• On 28 Jan 2011 at 08:18:05, wangchangguang (chg-wang.aaa.hotmail.com) sent the message
  

  
  Dear All,      I have big Nonspecific binding issues on one compound plasma protein  binding assay using ultrafiltration centrifugee devices. Recentely , I  both detect the apical(C1,V1) and bottom(C2,V2)liquid concentration  after spin, and also estimate the volume of both side.Can I use this  equation to calculate the PB%=100*(C1V1-C2V1)/(C1V1+C2V2) if the  recovery is acceptable(around 70%). any suggestion? Thanks !
  chang
   Department of Biochemistry
  UT Southwestern Medical Center
  5323 Harry Hines Boulevard
  Dallas, Texas 75390-9038
  
  

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• On 28 Jan 2011 at 11:30:27, rahul vats (rahulvats1983.at.gmail.com) sent the message
  

  
  Dear Sir,
   I would rather suggest you to go for ultracentrifugation method or RED  (rapid equillibrium device) method. If the nonspecific binding exceeds  5%, ultra filtration is not recommended for the determination of plasma  protein binding.
   Hope this will help
  
  

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• On 28 Jan 2011 at 08:39:05, Marjorie Cotler (msgcot2.at.optimum.net) sent the message
  

  
  The following message was posted to: PharmPK
  
  Please using the ultrafiltration leads to many examples of non-specific  binding.  My advice is to use the 96 well Harvard Plates, typical  recovery's are above 95% without having to resort to any correction  factors.
                                            Stanley Cotler
  
  

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• On 28 Jan 2011 at 21:22:06, vijaya bhargava (vijayabhargava.k.-a-.gmail.com) sent the message
  

  
  The following message was posted to: PharmPK
  
  also we can use the HT Dialysis approach for the determination of the
  plasma protein binding. RED inserts also serve the best purpose. But
  before doing any plasma protein binding, its better we go for
  checking the nonspecific binding to apparatus...
  hope this help you.
  Thanking you and regards,
  vijayabhargava.k
  
  

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• On 30 Jan 2011 at 19:11:51, "Tausif Ahmed (Dr.)" (tausif.a.-a-.saiadvantium.com) sent the message
  

  
  The following message was posted to: PharmPK
  
  Dear all,
  
  We have been regularly using rapid equilibrium device (RED) from thermo  for carrying out plasma protein binding assays and found the results  pretty consistent. As already suggested by other group members, if you  face problems of non specific binding, then ultra filtration is not the  recommended method.
  
  Thanks
  Tausif
  
  Tausif Ahmed, Ph.D.
  Associate Director, DMPK and Toxicology,
  Sai Advantium Pharma Ltd., Building 1, Plot No. 2, Chrysalis Enclave, International Biotech Park,
  Phase II, Hinjewadi, Pune - 411 057,
  Maharashtra, India
  
  

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