Fluorogenic dyes
AssigneeChiron Corporation
InventorsDaniel J. Suich, Ronald N. Zuckermann
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Abstract
The present invention relates to the use of fluorogenic or chromogenic dyes as reporter molecules for detecting cell entry by a specific molecule.
Claims28
A high throughput screening method for identifying compounds in a compound library that are transport compounds, wherein a transport compound refers to a compound that is internalized by a cell comprising:
- a. attaching the individual compounds in a compound library to a solid phase via a cleavable linkage, wherein each of said individual compounds is attached directly or indirectly to a fluorogenic or chromogenic reporter that becomes fluorescent or undergoes a change in light absorption properties upon cell entry;
- b. releasing the reporter-compound conjugate from the solid phase;
- c. contacting the released reporter-compound conjugates with a cell;
- and
- d. determining whether said compound is a transporter molecule based on whether there is a detectable change in fluorescence or light absorption.
A method for determining whether a compound is a transport molecule that is internalized by a cell comprising:
- a. contacting a cell with a conjugate comprising bis-acylated fluorescein or dichlorodithio-fluorescein disulfide reporter compound that only generates fluorescence after cell entry, which reporter is covalently bound to a potential transport compound which is to be screened for its ability to be internalized by the contacted cell;
- b. detecting whether fluorescence is generated after said contacting step;
- and
- c. correlating the generation of fluorescence to the internalization of the transport compound by the cell.
A library of compounds that is to be screened for their ability to be internalized by a desired cell that comprises a plurality of different compounds, wherein the different compounds in such library are covalently attached to a fluorogenic reporter molecule that becomes fluorescent after it internalizes a cell.
Dependencies are inferred from the claim text.