# US 10,371,703 B2 · Antibodies that inhibit long interspersed element-1 retrotransposon endonuclease activity

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- Number: US 10,371,703 B2
- Document ID: US10371703B2
- Status: Granted
- Kind: B2
- Application number: 14/943,942
- Filed: 2015-11-17
- Published: 2016-09-08
- Granted: 2019-08-06
- Earliest claimed: 2014-11-18
- Applicants: Victoria Perepelitsa Belancio; Mark Sokolowski
- Inventors: Victoria Perepelitsa Belancio; Mark Sokolowski
- Web page: https://patentgrep.com/patent/US10371703B2

## CPC

- G01N 33/573: for enzymes or isoenzymes
- C07K 16/40: against enzymes
- G01N 33/6863: {Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors}
- C07K 2317/33: Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C12Y 301/21: Endodeoxyribonucleases producing 5'-phosphomonoesters (3.1.21)
- G01N 2333/922: Ribonucleases (RNAses); Deoxyribonucleases (DNAses)
- 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)
- C12Y: ENZYMES

## Abstract

The present disclosure relates to monoclonal anti-LINE-1 ORF2 protein endonuclease domain antibody, and fragments and derivatives that bind the LINE-1 ORF2 protein endonuclease domain and inhibit its activity.

## Claims

### Claim 1

A binding reagent that specifically binds to human long interspersed element-1 (L1) open reading frame 2 (ORF2) protein having an endonuclease (EN) domain without an inactivating mutation at position 205, which binding reagent is selected from the group consisting of (a) an antibody having six complementarity determining regions (CDRs), which CDRs are the same as the six CDRs of monoclonal anti-L1 ORF2 protein EN antibody (hORF2 239), (b) a fragment of said antibody, said fragment having six CDRs which are the same as the six CDRs of monoclonal anti-L1 ORF2 protein EN antibody (hORF2 239) and which retains L1 ORF2 protein EN binding activity, and (c) a derivative of said antibody, said derivative having six CDRs which are the same as the six CDRs of monoclonal anti-L1 ORF2 protein EN antibody (hORF2 239), and which retains L1 ORF2 protein EN binding activity.

### Claim 2 (depends on 1)

The binding reagent of claim 1, wherein said derivative of said antibody is a chimeric or a humanized antibody.

### Claim 3 (depends on 1)

The binding reagent of claim 1, wherein said fragment is selected from the group consisting of a Fab, a Fab′, a F(ab′)2, a Fv, a helix-stabilized antibody, a diabody, a single-chain Fv (“scFv”), a disulfide stabilized antibody (“dsFv”), a domain antibody (“dAb,”) and a minibody.

### Claim 4 (depends on 1)

The binding reagent of claim 1, wherein said antibody is monoclonal anti-L1 ORF2 protein EN antibody (hORF2 239) or a fragment of said antibody which retains L1 ORF2 protein EN binding activity.

### Claim 5 (depends on 4)

The binding reagent of claim 4, wherein said antibody is monoclonal anti-L1 ORF2 protein EN antibody (hORF2 239).

### Claim 6 (depends on 4)

The binding reagent of claim 4, wherein said fragment is selected from the group consisting of a Fab, a Fab′, a F(ab′)2, a Fv; a helix-stabilized antibody, a diabody, a single-chain Fv (“scFv”), a disulfide stabilized antibody (“dsFv”), a domain antibody (“dAb,”) and a minibody.

### Claim 7 (depends on 1)

The binding reagent of claim 1, further wherein said binding reagent is labeled with a detectable label.

### Claim 8 (depends on 7)

The binding reagent of claim 7, further wherein label is conjugated to said binding reagent.

### Claim 9 (depends on 7)

The binding reagent of claim 7, further wherein label is fused to said binding reagent.

### Claim 10

A hybridoma producing a monoclonal antibody which is monoclonal anti-L1 ORF2 protein EN antibody (hORF2 239).

### Claim 11 (depends on 10)

The hybridoma of claim 10, which hybridoma is HORF2 hybridoma.

### Claim 12

A method of detecting whether human long interspersed element-1 (L1) open reading frame 2 (ORF2) protein having an endonuclease (EN) domain without an inactivating mutation at position 205 is present in a sample, said method comprising of contacting said sample with a binding regent which specifically binds to human L1ORF2 protein with said EN domain, which binding reagent is selected from the group consisting of (a) an antibody having six complementarity determining regions (CDRs), which CDRs are the same as the six CDRs of monoclonal anti-L1 ORF2 protein EN domain antibody (hORF2 239), (b) a fragment of said antibody having six CDRs, which CDRs are the same as the six CDRs of monoclonal anti-L1 ORF2 protein EN domain antibody (hORF2 239), and which retains L1 ORF2 protein EN domain binding activity, and (c) a derivative of said antibody having six CDRs, which CDRs are the same as the six CDRs of monoclonal anti-L1 ORF2 protein EN domain antibody (hORF2 239), and which retains L1 ORF2 protein EN domain binding activity, under conditions which permit binding of the binding reagent to said protein in said sample, and detecting whether any binding reagent has bound to said protein in said sample.

### Claim 13 (depends on 12)

The method of claim 12, further wherein said detection is by enzyme-linked immunosorbent assay (“ELISA”).

### Claim 14 (depends on 12)

The method of claim 12, further wherein said antibody is a monoclonal antibody.

### Claim 15 (depends on 14)

The method of claim 14, wherein said antibody is monoclonal anti-L1 ORF2 protein EN antibody (hORF2 239) or a fragment said antibody which retains L1 ORF2 protein EN domain binding activity.

### Claim 16 (depends on 15)

The method of claim 15, wherein said fragment is selected from the group consisting of a Fab, a Fab′, a F(ab′)2, a Fv; a helix-stabilized antibody, a diabody, a single-chain Fv (“scFv”), a disulfide stabilized antibody (“dsFv”), a domain antibody (“dAb,”) and a minibody.

### Claim 17 (depends on 12)

The method of claim 12, further wherein said binding reagent is labeled with a detectable label.

### Claim 18 (depends on 12)

The method of claim 12, further wherein label is conjugated to said binding reagent.