C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/02 (2006.01) C12M 1/34 (2006.01) C12Q 1/68 (2006.01) G01N 33/50 (2006.01) G01N 35/02 (2006.01) G01N 35/04 (2006.01) G01N 35/10 (2006.01)
Patent
CA 2121506
Highly efficient automated measurement apparatus and methods for automated drug screening procedures are provided. The apparatus is capable of initiating and measuring rapid or transient events such as cell receptor and/or ion channel activity. The apparatus of the invention is capable of aligning with predetermined positions, one or more samples contained in a multi- well container, initiating the reaction with reagent addition and measuring a resultant attribute for a period of time. The appara- tus is capable of substantially continuously measuring and recording data corresponding to the measured attribute before, during and after initiation of the reaction so that a time course of the rapid or transient event may be determined. After the reaction(s) are complete in one or more wells of the multi-well container, the apparatus can align one or more different wells to be assayed with the predetermined position and repeat the cycle until a predetermined number of wells are assayed. Automated drug screen- ing methods employing the apparatus are also provided for screening for compounds which activate, inhibit or potentiate cellular ion channel or receptor activity. The assays are fluorescent indicator-based assays which utilize viable cells containing in their cyt- oplasm effective levels of a fluorescent indicator which is responsive to changes in ion concentration. Activation of the ion chan- nels or receptors is initiated in the automated measurement apparatus by injection into one or more predetermined cell-contain- ing wells of a reagent which is a known or putative activator of the ion channels or receptors of interest. The resultant activity of the ion channels or receptors (which causes changes in ion concentration in the cytoplasm) is determined by measurement of flu- orescence intensity changes of the indicator in response to an excitation wavelength.
Akong Michael Anthony
Brust Paul
Harpold Michael Miller
Velicelebi Gonul
Merck Sharp & Dohme Corp.
Smart & Biggar
The Salk Institute Biotechnology/industrial Associates Inc.
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