# US 5,601,992 A · Peptide library formats and methods relating thereto

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- Number: US 5,601,992 A
- Document ID: US5601992A
- Status: Granted
- Kind: A
- Application number: 08/303,585
- Filed: 1994-09-09
- Granted: 1997-02-11
- Earliest claimed: 1933-05-28
- Assignees: Bunsen Rush Laboratories, Inc.
- Inventors: Michael R. Lerner; Channa K. Jayawickreme; Ethan A. Lerner
- Web page: https://patentgrep.com/patent/US5601992A

## CPC

- C40B 30/04: by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
- G01N 33/531: Production of immunochemical test materials
- G01N 33/54353: {with ligand attached to the carrier via a chemical coupling agent (coatings G01N33/54393)}
- G01N 33/6845: {Methods of identifying protein-protein interactions in protein mixtures}
- G01N 33/6878: {in epitope analysis}
- G01N 2333/5758: Gastrin releasing peptide
- G01N 2410/00: Assays, e.g. immunoassays or enzyme assays, involving peptides of less than 20 animo acids
- C40B: COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES (in silico combinatorial libraries of nucleic acids, proteins or peptides G16B35/00; in silico combinatorial chemistry G16C20/60)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A method for detecting the interaction between an oligomeric molecule and a target is provided. A plurality of beads is applied to a substrate, each bead having associated therewith multiple copies of only a single oligomeric molecule. The beads are substantially spaced-apart from one another and are substantially immobilized on the substrate. Conditions are applied such that a substantial portion of the multiple copies associated with each bead can disassociate from and diffuse into the substrate, the substrate constructed and arranged to permit only substantially localized diffusion. A localized signal occurring as a result of the diffusion is detected. Coated beads and libraries of coated beads also are provided.

## Claims

### Claim 1

A method for detecting the interaction between a peptide and a target, comprising:
(I) placing the target in a container;
(ii) overlaying the target with a substrate that is free of the target, the substrate constructed and arranged to permit only substantially localized diffusion of the peptides therein;
(iii) applying a plurality of beads to the surface of the substrate in a manner such that the beads are separated from the target by the substrate and such that the beads are substantially spaced-apart from one another and are immobilized on the substrate, each bead having associated therewith multiple copies of only a single peptide, wherein at least a substantial portion of the multiple copies is associated with each bead via non-covalent bonds;
(iv) allowing the substantial portion of the multiple copies to diffuse through the substrate and interact with the target, thereby resulting in a localized signal at the target;
(v) detecting the localized signal; and
(vi) associating the localized signal with one or a limited number of the plurality of beads applied to the substrate.

### Claim 2 (depends on 1)

The method of claim 1, further comprising
(vii) isolating a bead in the vicinity of the localized signal; and
(viii) sequencing the peptide associated with the bead in the vicinity of the localized signal.

### Claim 3 (depends on 1)

The method of claim 1, wherein the target is a living cell.

### Claim 4

A method for detecting the interaction between a peptide and a target comprising:
(I) attaching linkers to a plurality of beads;
(ii) attaching a plurality of peptides to the beads via covalent bonds to the linkers, each bead having associated therewith multiple copies of only a single peptide;
(iii) cleaving at least some of the covalent bonds so that at least a substantial portion of the multiple copies are associated with each bead via noncovalent bonds;
(iv) placing the target in a container;
(v) overlaying the target with a substrate that is free of the target, the substrate constructed and arranged to permit only localized diffusion of the peptides therein;
(vi) applying the plurality of beads to the substrate in a manner such that the beads are separated from the target by the substrate and such that the beads are substantially spaced-apart from one another and are immobilized on the substrate;
(vii) allowing the substantial portion of the multiple copies to diffuse through the substrate and interact with the target, thereby resulting in a localized signal at the target;
(vii) detecting the localized signal;
(ix) associating the localized signal with one or a limited number of the plurality of beads applied to the substrate;
(x) isolating a bead in the vicinity of the localized signal;
(xi) sequencing the peptide at the bead in the vicinity of the localized signal.

### Claim 5

A method for determining the relative potency of an interaction between a peptide and a target, the method comprising:
(I) placing the target in a container;
(ii) overlaying the target with a substrate that is free of the target, the substrate constructed and arranged to permit only localized diffusion of the peptides therein;
(iii) applying a plurality of beads to the substrate in a manner such that the beads are separated from the target by the substrate and such that the beads are substantially spaced-apart from one another and are immobilized on the substrate, each bead having associated therewith copies of only a single peptide, wherein at least a substantial portion of the multiple copies is associated with each bead via non-covalent bonds;
(iv) allowing the substantial portion of the multiple copies to diffuse through the substrate and interact with the target, thereby resulting in a localized signal associated with the target;
(v) associating the localized signal with one or a limited number of the plurality of beads applied to the substrate;
(vi) measuring the localized signal at pre-selected time intervals; and
(vii) determining the rate of change of the localized signal as a function of the time intervals, whereby the rate of change is proportional to the potency of the interaction between the peptide and the target.