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Hi Amol,
Attached you can find the review. [Sorry attachments aren't possible
with PharmPK - db]
It seems to me that your NCE binds to the filter unit (or precipitates
in water, which is only visible from say 30 ug/ml). Non specific
binding is unfavourable, but if it is constant of a concentration
range, one could work with it.
Also I would recommend to incubate your NCE with the plasma (and of
course also in water, preferable PBS actually) at least 1 hr at 37=B0C
to reach equilibrium (I don't know if you did, it was not in the mail).
Then you should try to keep separation between bound and free (the
centrifugation step) as short as possible and at the right temperature
(which you allready do). This is because the separation influences the
equilibrium, so filter just enough to be able to measure. Temperature
of course also influences the equilibrium.
Hoped to give you some useful tips,
sylvia
Sylvia Lemstra
Research Scientist
Pharmacology & preclinical development
4SC AG tel: +49 89 700 763-0
Am Klopferspitz 19a fax: +49 89 700 763-29
82152 Martinsried e-mail: lemstra.-at-.4sc.com
Germany Internet www.4sc.com
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