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US 6,087,103 AGranted
Tagged ligand arrays for identifying target-ligand interactions
AssigneeLifeSpan BioSciences, Inc.
InventorGlenna C. Burmer
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Abstract
The present invention relates generally to high throughput screening methods. More particularly, the present invention provides screening methods that can readily be used to identify simultaneously multiple proteins or compounds that interact with multiple ligands, using a tagged array of ligands.
Claims31
A method of screening a target for ligand binding, the method comprising the steps of:
- a. providing a library of tags, each tag having a known tag address that is located by reference to a matrix;
- b. providing a library of ligands, each ligand having a known ligand address that is located by reference to a matrix;
- c. binding a tag having a known address to each ligand to create tagged ligands, wherein the ligand address corresponds to the tag address;
- d. incubating a target with at least two tagged ligands;
- e. determining whether or not at least one tagged ligand binds to a target;
- and
- f. identifying said at least one tagged ligand bound to the target by identifying the tag having a known address bound to said at least one tagged ligand, wherein the tag address indicates the corresponding ligand address.
4 more elements
A method of simultaneously screening target proteins for ligand binding, the method comprising the steps of:
- a. providing a library of recombinant oligonucleotides, each oligonucleotide having a known tag address that is located by reference to a matrix;
- b. providing an expressed cDNA library of ligand proteins, each ligand protein having a known ligand address that is located by reference to a matrix;
- c. binding each ligand protein to an oligonucleotide having a known tag address to create tagged ligand proteins, wherein the ligand address corresponds to the tag address;
- d. pooling the tagged ligand proteins;
- e. incubating an expressed cDNA library of target proteins with the pooled tagged ligand proteins;
- f. determining whether or not at least one tagged ligand protein binds to a target protein;
- g. amplifying the oligonucleotide from said at least one tagged ligand protein bound to the target;
- h. hybridizing the amplified oligonucleotide to an oligonucleotide bound to a membrane, wherein the membrane provides the matrix corresponding to the tag address of each oligonucleotide;
- and
- i. identifying said at least one tagged ligand protein bound to the target by identifying the oligonucleotide having a known tag address bound to said at least one tagged ligand protein, wherein the tag address indicates the corresponding ligand address.
7 more elements
A method of simultaneously screening for ligands expressed by a cancer cell that bind to targets expressed by a cancer cell, the method comprising the steps of:
- a. providing a library of tags, each tag having a known tag address that is located by reference to a matrix;
- b. providing an expressed cancer cell cDNA library of ligand proteins, each ligand protein having a known ligand address that is located by reference to a matrix;
- c. binding a tag having a known address to each ligand protein to create tagged ligand proteins, wherein the ligand address corresponds to the tag address;
- d. expressing target proteins from the cancer cell cDNA;
- e. incubating the target proteins with the tagged ligand proteins;
- f. determining whether or not a tagged ligand protein binds to a target protein;
- and
- g. identifying the tagged ligand protein bound to the target by identifying the tag.
5 more elements
An array of microtiter plates, comprising:
- a. a first plate comprising wells containing a library of tags, wherein each well contains a unique tag, thereby providing each tag with a known tag address that is located by reference to a matrix;
- b. a second plate comprising wells containing a library of ligands, each ligand having a known ligand address that is located by reference to a matrix, wherein each ligand is bound to a tag having a known address, and wherein each well contains a unique ligand, thereby providing the ligand with an address corresponding to the tag address.
An array of microtiter plates, comprising:
- a. a first plate comprising wells containing a library of oligonucleotide tags, wherein each well contains a unique oligonucleotide tag, thereby providing each oligonucleotide tag with a known tag address that is located by reference to a matrix;
- b. a second plate comprising wells containing a library of ligands, each ligand having a known ligand address that is located by reference to a matrix, wherein each ligand is bound to an oligonucleotide tag having a known address, and wherein each well contains a unique ligand, thereby providing the ligand with an address corresponding to the tag address.
Dependencies are inferred from the claim text.