Pharmacopeia Inc.

Daniel Chelsky

Jonathan J Burbaum

Gregory L Kirk

James Inglese

Joseph J Brzezinski

Thomas Dy Chung

T C Ramaraj

Thirleen G Nichols

A homogeneous high throughput assay is described which screens compounds for enzyme inhibition, or receptor or other target binding. Inhibition (or binding) by the library compounds causes a change in the amount of an optically detectable label that is bound to suspendable cells or solid supports. The amounts of label bound to individual cells or solid supports are microscopically determined, and compared with the amount of label that is not bound to individual cells or solid supports. The degree of inhibition or binding is determined using this data. Confocal microscopy, and subsequent data analysis, allow the assay to be carried out without any separation step, and provide for high throughput screening of very small assay volumes using very small amounts of test compound.

High-throughput assay

A lawn assay is described for determining compounds that affect enzyme activity or that bind to target molecules. Compounds to be screened are cleaved, and diffused from solid supports into a colloidal matrix. Enzymatic catalysis or binding to target molecules by the compounds is carried out in the matrix. Active compounds are found by monitoring a photometrically detectable change in a substrate, coenzyme, or cofactor involved in the enzymatic reaction, or in a labeled ligand bound to the target molecule, that produces a zone of activity associated with the compounds.

Lawn assay for compounds that affect enzyme activity or bind to target

A system which precisely arrays small objects into a plurality of containers which comprises a vessel containing the objects in a fluid suspension; a tube, the proximal end of which is positioned substantially at the bottom of the vessel and the distal end of which is connected to a member which directs gravitationally-formed fluid drops from the distal end downward into a container positioned below said distal end; a small-objects detector, for detecting the objects in the fluid in the tube, disposed near the distal end; a drop detector connected to the member below the distal end; a device for controlling flow of the suspension through the tube by pressurizing the vessel; or device for discriminating between those signals from the small-objects detector caused by the small objects and those signals caused by other events, i.e., not caused by the small objects; a device for comparing signals from the small-objects detector caused by the small objects with signals from the drop detector; and a device for determining the number of the objects deposited into the container. The system further comprises a positioning device for re-positioning the distal end of the tube from one to another, with timing derived from the small-objects and drop detectors. The positioning device comprises an X-Y transport which is either 1) a movable member which moves the distal end of the tube and the detectors or 2) an X-Y stage which moves the containers, with the tube and the detectors attached to a stationary member.

Partitioning device

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Moylan, PA, United States of America

Daniel Chelsky