# US 8,569,043 B2 · Detection of post-translationally modified peptides with liquid crystals

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- Number: US 8,569,043 B2
- Document ID: US8569043B2
- Status: Granted
- Kind: B2
- Application number: 12/837,471
- Filed: 2010-07-15
- Published: 2011-03-03
- Granted: 2013-10-29
- Earliest claimed: 2003-09-23
- Assignees: Wisconsin Alumni Research Foundation
- Applicants: Nicholas L. Abbott; Brian H. Clare; Paul J. Bertics
- Inventors: Nicholas L. Abbott; Brian H. Clare; Paul J. Bertics
- Web page: https://patentgrep.com/patent/US8569043B2

## CPC

- G01N 33/542: with steric inhibition or signal modification, e.g. fluorescent quenching
- B01J 19/0046: {Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays (synthesis methods per se C40B50/00)}
- B82Y 15/00: Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
- B82Y 30/00: Nanotechnology for materials or surface science, e.g. nanocomposites
- B82Y 40/00: Manufacture or treatment of nanostructures
- G01N 33/54373: {involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings}
- G01N 33/6842: {Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins}
- B01J 2219/00382: Stamping
- B01J 2219/00596: Solid-phase processes
- B01J 2219/00659: Two-dimensional arrays
- B01J 2219/0072: Organic compounds
- B01J 2219/00722: Nucleotides
- B01J 2219/00725: Peptides
- B01J 2219/0074: Biological products
- B01J 2219/00743: Cells
- B01J 2219/00745: Inorganic compounds
- B01J 2219/0075: Metal based compounds
- C40B 40/06: Libraries containing nucleotides or polynucleotides, or derivatives thereof
- C40B 40/10: Libraries containing peptides or polypeptides, or derivatives thereof
- C40B 40/18: Libraries containing only inorganic compounds or inorganic materials
- C40B 60/14: for creating libraries
- 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)
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- 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)

## Abstract

A method for differentiating between a post-translationally modified peptide and a peptide contained in a sample, comprising: (a) contacting the sample with a peptide attachment surface to create a peptidized surface, wherein the sample includes at least one functional group; (b) contacting the peptidized surface with a recognition reagent that selectively binds or forms a complex with the post-translationally modified peptide in the sample to provide an incubated surface; and (c) contacting a liquid crystal with the incubated surface and detecting presence of post-translationally modified peptide in the sample with the liquid crystal.

## Claims

### Claim 1

A device for differentiating between a peptide and a post-translationally modified form of the peptide, comprising:
(a) a support having a peptide attachment surface;
(b) a peptide and a post-translationally modified form of the peptide are immobilized on the peptide attachment surface; and
(c) a liquid crystal in contact with the peptide and the post-translationally modified form of the peptide that are immobilized on the peptide attachment surface,
wherein the orientation of the liquid crystal in contact with the post-translationally modified form of the peptide is different than the orientation of the liquid crystal in contact with the peptide.

### Claim 2 (depends on 1)

The device of claim 1, wherein the post-translationally modified form of the peptide, the peptide, or both are thiol-containing or disulfide-containing peptides.

### Claim 3 (depends on 1)

The device of claim 1, wherein the post-translationally modified form of the peptide, the peptide, or both are thiolated peptides.

### Claim 4 (depends on 1)

The device of claim 1, wherein the peptide attachment surface is selected from the group consisting of glass, silica, silicon, gold, titania, titanium, alumina or aluminum.

### Claim 5 (depends on 4)

The device of claim 4, wherein the peptide attachment surface comprises a layer of gold.

### Claim 6 (depends on 4)

The device of claim 4 wherein the peptide attachment surface comprises a layer of titania.

### Claim 7 (depends on 5)

The device of claim 5, wherein the layer of gold overlies a layer of a material that promotes adhesion of the gold to the support.

### Claim 8 (depends on 6)

The device of claim 6, wherein the material that promotes adhesion of the gold to the support is titanium.

### Claim 9 (depends on 1)

The device of claim 1, wherein the post-translationally modified form of the peptide is selected from the group consisting of a phosphorylated peptide, an acylated peptide, a glycosylated peptide, an adenylated peptide, a farnesylated peptide, an alkylated peptide, a proteolytically cleaved peptide, and mixtures thereof.

### Claim 10 (depends on 1)

The device of claim 1, wherein the peptide attachment surface comprises a recognition reagent that selectively binds or forms a complex with the post-translationally modified peptide.

### Claim 11 (depends on 10)

The device of claim 10 wherein the recognition reagent comprises a phosphate-binding compound.

### Claim 12 (depends on 10)

The device of claim 10, wherein the recognition reagent is selected from the group consisting of an antibody or an antibody fragment, a cationic compound, a cationic surfactant, a polyelectrolyte, a cationic iron compound, a phosphosensor dye and mixtures thereof.

### Claim 13 (depends on 1)

The device of claim 1, wherein the liquid crystal is selected from the group consisting of nematic, smectic, polymeric, thermotropic, lyotropic, columnar, nematic discotic, calamitic nematic, ferroelectric, discoid, cholesteric and mixtures thereof.

### Claim 14 (depends on 1)

The device of claim 1, further comprising a second surface, wherein the second surface contacts the liquid crystal in contact with the peptide and the post-translationally modified form of the peptide.