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Background:
The pharmacokinetic and pharmacodynamic properties of drugs are influenced by the extent of binding to plasma proteins, as only unbound "free" drug has the ability to partition across membranes, undergo metabolism and clearance by the kidney, as well as interact with targets to exert an effect. 

Drugs generally bind reversibly to plasma proteins (mostly albumin, a1-acid glycoprotein and lipoproteins) with different kinetics and affinity.  Highly protein bound drugs are retained in the plasma compartment, with restricted distribution into tissues, decreased metabolism, clearance (low extraction ratio drugs), prolonged half-lives and limited brain penetration. 

The degree to which a compound binds to plasma proteins is determined across species during early drug discovery. Equilibrium dialysis is a commonly used technique to measure plasma protein binding (PPB).
  

   Assay description

   Technique
      equilibrium dialysis

   Species
      mouse, rat , human, dog,  rabbit,  monkey

   Compound concentration
      5µM (0.5% DMSO)

   Compound requirements
      50µl of 10mM stock solution or
      1-2 mg of dry matter

   Detection method
      LC-MS/MS with internal standard

   Results
      %fraction bound (%Fb)
      recovery



Figure 1. %Fb values obtained for
reference compounds in 4 different species

 

 


Figure 2. %Fb values of commercial compounds obtained
in human plasma in 3 separate experiments

Assay controls
   protein contamination check
   reference compounds:  acebutolol, propranolol, verapamil and nicardipine  (Figure 1)
   data obtained for 10 commercial  compounds (Figure 2)

Assay details adjustable to client’s and/or project specific requests

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