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Hi, can someone provide specific information regarding how to minimize
nonspecific protein binding in plasma protein binding studies. I am
interested in the techniques that might be use e.g. using TWEEN or
some kind of equillibrium dialysis cells where nonspecific binding is
minimal.
Regards.
Neil
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Hi Neil,
Nonspecific binding can be minimized by using Ultracentrifugation
technique, since bound & unbound fractions are separated on the basis
of their sedimentation coefficient & there is no interaction with any
of the membrane used in other two techniques Ultrafiltration &
Equilibrium dialysis.
Regards
Sachin Latad
Dept. of ADME, Discovery Research
Torrent Research Centre
Near Indira Bridge,
P.O. BHAT - 382 428
Dist. Gandhinagar (Gujarat, India)
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The following message was posted to: PharmPK
Dear Neil:
We have had the pleasure of working with a series of very lipophilic
compounds that exhibit extensive non-specific binding. We tried many
different variations of equilibrium dialysis but to no avail. Recently
we tried an approach published by Taylor and Harker using centrifugal
ultra-filtration and have been able to routinely achieve >95% recovery
of compounds. The reference is
Taylor S, Harker A. J. Pharm. Biomed. Anal. 2006 Apr 11;41(1):299-303.
Modification of the ultrafiltration technique to overcome solubility and
non-specific binding challenges associated with the measurement of
plasma protein binding of corticosteroids.
Best wishes,
Damon
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The following message was posted to: PharmPK
Hi Neil,
Non-specific binding can be minimized by using Blood partitioning
technique,
which partitions the drug between the red blood cells membrane and
plasma
proteins. While working with increasing protein concentration you can
determine the binding coefficient: NKa)
Hopping this can help you
Regards
Francoise BREE
XENOBLIS
Parc d'Affaires de la Breteche
Batiment B1
35760 Saint Gregoire
France
francoise.bree.at.xenoblis.com
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Dear all,
I'm Rima Sturm, I'm working in Basel for a pharmaceutical company, and
I saw in the "PharmPK Discussion" on internet that you're searching a
new method in order to minimize the nonspecific protein binding in
plasma protein binding studies.
I found a very interesting publication: "Modulation of Nonspecific
Binding in Ultra filtration Protein Binding Studies" in The
Pharmaceutical Research Journal (Vol. 20, No. 7, July 2003). I tried
to do the same experiment as the one described in this previous
publication but my results are not good. I have the impression that
the Tween80 block the filter because my recovery is not good at all!
Did you already try this technique? Do you have an idea, how to wash
out good this Tween80?
Thank you very much for your answer.
Best regards,
Rima Sturm
--
Rima Sturm
Pharmacology
email: rima.sturm.-at-.basilea.com
--
Basilea Pharmaceutica International Ltd.
Grenzacherstrasse 487
Postfach
CH-4005 Basel
Switzerland
www.basilea.com
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