# US 9,040,310 B2 · Antibody-nanoparticle conjugates and methods for making and using such conjugates

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- Number: US 9,040,310 B2
- Document ID: US9040310B2
- Status: Granted
- Kind: B2
- Application number: 13/640,944
- Filed: 2011-04-27
- Published: 2013-02-07
- Granted: 2015-05-26
- Earliest claimed: 2010-04-27
- Assignees: Ventana Medical Systems, Inc.
- Applicants: Julia Ashworth-Sharpe; Chol Steven Yun; Zhanna Zhilina; Adrian E. Murillo; Donald D. Johnson; Michael Farrell; Jerome W. Kosmeder; Christopher Bieniarz
- Inventors: Julia Ashworth-Sharpe; Chol Steven Yun; Zhanna Zhilina; Adrian E. Murillo; Donald D. Johnson; Michael Farrell; Jerome W. Kosmeder; Christopher Bieniarz
- Web page: https://patentgrep.com/patent/US9040310B2

## CPC

- G01N 33/54346: {Nanoparticles}
- C07F 9/12: with hydroxyaryl compounds
- C07F 9/5004: {Acyclic saturated phosphines}
- C07F 9/5022: {Aromatic phosphines (P-C aromatic linkage)}
- C07F 9/5045: {Complexes or chelates of phosphines with metallic compounds or metals}
- C07F 9/5728: {condensed with carbocyclic rings or carbocyclic ring systems}
- C07F 9/65515: {the oxygen atom being part of a five-membered ring}
- C07F 9/65517: {condensed with carbocyclic rings or carbocyclic ring systems}
- C07F 9/65522: {condensed with carbocyclic rings or carbocyclic ring systems}
- G01N 33/531: Production of immunochemical test materials
- G01N 33/553: Metal or metal coated
- G01N 33/587: {Nanoparticles}
- G01N 33/6854: {Immunoglobulins}
- G01N 2333/916: acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
- 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)
- C07F: ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM (metal-containing porphyrins C07D487/22)

## Abstract

Disclosed herein are antibody-nanoparticle conjugates that include two or more nanoparticles (such as gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof) directly linked to an antibody or fragment thereof through a metal-thiol bond. Methods of making the antibody-nanoparticle conjugates disclosed herein include reacting an arylphosphine-nanoparticle composite with a reduced antibody to produce an antibody-nanoparticle conjugate. Also disclosed herein are methods for detecting a target molecule in a sample that include using an antibody-nanoparticle conjugate (such as the antibody-nanoparticle conjugates described herein) and kits for detecting target molecules utilizing the methods disclosed herein.

## Claims

### Claim 1

An antibody-nanoparticle conjugate, comprising three or more metal nanoparticles directly linked to a single antibody through three or more metal-thiol bonds,
wherein the metal-thiol bonds are between thiol from of cysteine residues of the antibody and the three or more metal nanoparticle without an intervening linker.

### Claim 2 (depends on 1)

The antibody-nanoparticle conjugate of claim 1, wherein the conjugate comprises three to seven nanoparticles.

### Claim 3 (depends on 2)

The antibody-nanoparticle conjugate of claim 2, wherein the conjugate comprises five nanoparticles.

### Claim 4 (depends on 1)

The antibody-nanoparticle conjugate of claim 1, wherein the antibody comprises a rabbit, goat, mouse, or anti-hapten antibody.

### Claim 5 (depends on 1)

The antibody-nanoparticle conjugate of claim 1, wherein the nanoparticle comprises gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof.

### Claim 6 (depends on 1)

The antibody-nanoparticle conjugate of claim 1, wherein the nanoparticle is about 5 nm or less in diameter.

### Claim 7 (depends on 6)

The antibody-nanoparticle conjugate of claim 6, wherein the nanoparticle is about 0.5 to 5 nm in diameter.