# US 11,112,413 B2 · Multiplexed immunohistochemistry using recombinant antibodies with epitope tags

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- Number: US 11,112,413 B2
- Document ID: US11112413B2
- Status: Granted
- Kind: B2
- Application number: 16/123,750
- Filed: 2018-09-06
- Published: 2019-03-28
- Granted: 2021-09-07
- Earliest claimed: 2016-03-08
- Assignees: Ventana Medical Systems, Inc.
- Applicants: Ventana Medical Systems, Inc.
- Inventors: Michael Farrell; Antony Hubbard; Eric May; Lei Tang; Tsu-Shuen Tsao; Wenjun Zhang
- Web page: https://patentgrep.com/patent/US11112413B2

## CPC

- G01N 33/6878: {in epitope analysis}
- C07K 16/18: against material from animals or humans
- C07K 16/2809: {against the T-cell receptor (TcR)-CD3 complex}
- C07K 16/2815: {against CD8}
- C07K 16/2827: {against B7 molecules, e.g. CD80, CD86}
- C07K 16/2887: {against CD20}
- C07K 16/2896: {against molecules with a "CD"-designation, not provided for elsewhere}
- C07K 16/32: against translation products of oncogenes
- C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation
- C07K 2319/42: containing a HA(hemagglutinin)-tag
- G01N 2570/00: Omics, e.g. proteomics, glycomics or lipidomics; Methods of analysis focusing on the entire complement of classes of biological molecules or subsets thereof, i.e. focusing on proteomes, glycomes or lipidomes
- 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)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The present disclosure is directed to epitope-tagged antibodies, as well as methods of employing the epitope-tagged antibodies for detecting one or more targets in a biological sample, e.g. a tissue sample.

## Claims

### Claim 1

A kit comprising:
(a) at least one epitope-tagged antibody, wherein the at least one epitope-tagged antibody comprises an antibody and at least one epitope tag construct, and
(b) detection reagents for detecting the at least one epitope-tagged antibody,
wherein the detection reagents comprise at least one anti-tag antibody, wherein the at least one anti-tag antibody is specific for an expressed epitope tag of the at least one epitope-tagged antibody, and
wherein the at least one epitope tag construct consists of the amino acid sequence selected from the group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, and SEQ ID NO:32.

### Claim 2 (depends on 1)

The kit of claim 1, wherein the at least one anti-tag antibody comprises a fluorophore.

### Claim 3 (depends on 1)

The kit of claim 1, wherein the at least one epitope-tagged antibody comprises at least one epitope-tagged antibody specific for FoxP3, at least one epitope-tagged antibody specific for CD8, at least one epitope-tagged antibody specific for CD68, at least one epitope-tagged antibody specific for CD3, and at least one epitope-tagged antibody specific for CD20.

### Claim 4 (depends on 1)

The kit of claim 1, wherein the at least one epitope tag construct is expressed at a terminal end of a heavy chain antibody.

### Claim 5 (depends on 1)

The kit of claim 1, wherein the at least one epitope-tagged antibody is specific for the antigen human FoxP3.

### Claim 6 (depends on 5)

The kit of claim 5, wherein the at least one epitope tag construct consists of the amino acid sequence of SEQ ID NO:22.

### Claim 7 (depends on 1)

The kit of claim 1, further comprising instructions for use.