# US 6,720,190 B1 · Methods for screening compound libraries

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- Number: US 6,720,190 B1
- Document ID: US6720190B1
- Status: Granted
- Kind: B1
- Application number: 09/276,444
- Filed: 1999-03-25
- Granted: 2004-04-13
- Earliest claimed: 1998-03-27
- Inventors: Ole Hindsgaul; David C. Schriemer
- Web page: https://patentgrep.com/patent/US6720190B1

## CPC

- G01N 33/68: involving proteins, peptides or amino acids {(involving lipoproteins G01N33/92)}
- G01N 33/538: by sorbent column, particles or resin strip {, i.e. sorbent materials}
- G01N 33/54306: {Solid-phase reaction mechanisms}
- G01N 33/54366: {Apparatus specially adapted for solid-phase testing}
- G01N 33/6845: {Methods of identifying protein-protein interactions in protein mixtures}
- G01N 33/6848: {Methods of protein analysis involving mass spectrometry}
- H01J 49/0036: {Step by step routines describing the handling of the data generated during a measurement}
- B01J 2219/00585: Parallel processes
- B01J 2219/00704: integrated with the reactor apparatus
- B01J 2219/00707: separated from the reactor apparatus
- B01J 2219/00731: Saccharides
- B01J 2219/00738: Organic catalysts
- B01J 2219/00745: Inorganic compounds
- B01J 2219/00747: Catalysts
- C40B 30/08: by measuring catalytic activity
- C40B 40/12: Libraries containing saccharides or polysaccharides, or derivatives thereof
- C40B 40/18: Libraries containing only inorganic compounds or inorganic materials
- G01N 30/466: {with separation columns in parallel}
- G01N 30/7266: {by electric field, e.g. electrospray}
- G01N 2030/628: {Multiplexing, i.e. several columns sharing a single detector}
- H01J 49/00: Particle spectrometers or separator tubes
- Y10T 436/24: Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry
- 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)
- H01J: ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS (spark-gaps H01T; electric arc lamps H05B31/00; particle accelerators H05H)
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- 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)
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

Disclosed are methods for screening compound libraries using frontal chromatography in combination with mass spectrometry to identify and rank those members of the library that bind to a target receptor. Methods are also disclosed which permit a compound library to be rapidly screened to determine if any member of the library has an affinity for the target receptor as measured by a pre-selected indicator compound.

## Claims

### Claim 1

A method for screening a compound library to determine the relative affinity of a plurality of putative ligands to a target receptor or a plurality of target receptors relative to an indicator compound or a plurality of indicator compounds, which method comprises:
(a) providing a compound library comprising a plurality of putative ligands;
(b) providing at least one void marker compound;
(c) providing an indicator compound or a plurality of indicator compounds for each target receptor, each indicator compound having a pre-determined affinity for the target receptor and having a pre-determined break through time on the column in the absence of the compound library relative to a void marker compound;
(d) applying the compound library to a column comprising a target receptor or a plurality of target receptors, each target receptor selected from the group consisting of proteins, peptides, DNA, DNA fragments, RNA and RNA fragments and optionally bound to a solid phase support, under frontal chromatography conditions to partially equilibrate the column with the compound library;
(e) applying (i) a mixture comprising the compound library, the void marker compound and the indicator compound or compounds, or (ii) the void marker compound and the indicator compound or compounds, to the column under frontal chromatography conditions to provide an effluent;
(f) analyzing the effluent to determine a break through time for the indicator compound or compounds; and
(g) determining whether any putative ligands of the compound library have an affinity for a target receptor greater than the indicator compound by comparing the break through time for the indicator compound from step (f) with the predetermined break through time for the indicator compound in the absence of the compound library.

### Claim 2 (depends on 1)

The method of claim 1, wherein the analysis of the effluent is conducted using a mass spectrometer.

### Claim 3 (depends on 2)

The method of claim 2, wherein the mass spectrometer is an electrospray mass spectrometer.

### Claim 4 (depends on 1)

The method of claim 1, wherein the compound library comprises less than about 50,000 putative ligands.

### Claim 5 (depends on 4)

The method of claim 4, wherein the compound library comprises about 5 to about 100 putative ligands.

### Claim 6 (depends on 1)

The method of claim 1, wherein the compound library comprises putative ligands selected from the group consisting of carbohydrates, monosaccharides, oligosaccharides, polysaccharides, amino acids, peptides, oligopeptides, polypeptides, proteins, nucleosides, nucleotides, oligonucleotides, polynucleotides, lipids, retinoids, steroids, glycopeptides, glycoproteins, glycolipids, and proteoglycans.

### Claim 7 (depends on 1)

The method of claim 1, wherein the compound library comprises putative ligands selected from the group consisting of natural products.

### Claim 8 (depends on 1)

The method of claim 1, wherein the compound library comprises putative ligands selected from the group consisting of synthetic small molecule organic compounds.

### Claim 9 (depends on 1)

The method of claim 1, wherein each target receptor is bound to a solid phase support.

### Claim 10 (depends on 1)

The method of claim 1, wherein the column contains from about 1 fmol to about 10 nmol of target receptor active sites.

### Claim 11 (depends on 1)

The method of claim 1, wherein the pre-determined break through time for the indicator compound in the absence of the compound library is less than about 5 minutes.