Anti-acetaminophen antibodies and acetaminophen protein adducts

• US 10,570,216 B2
• Filed: 12/03/2015
• Issued: 02/25/2020
• Est. Priority Date: 12/03/2014
• Status: Active Grant

• Current Assignee: Arkansas Children'S Research Institute, Bioventures, LLC
• Original Assignee: Bioventures, LLC
• Inventors: Roberts, Dean W., James, Laura, Hinson, Jack
• Primary Examiner(s): Haq, Shafiqul
• Application Number:
  US15/532,418
• Publication Number:
  US 20170362340A1
• Time in Patent Office: 1,545 Days
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 19/00 Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/9486 Analgesics, e.g. opiates, aspirine

Abstract

The present disclosure provides isolated antibodies that bind to acetaminophen-protein adducts that are useful in the detection and diagnosis of acetaminophen-induced toxicity.

Claims

1. An isolated antibody, wherein the antibody specifically binds an acetaminophen-protein adduct but does not specifically bind free acetaminophen and recognizes the immunogen:

Carrier Protein-2-iminothiolane-APAP, wherein the antibody comprises;

(a) a light chain CDR1 amino acid sequence of SEQ ID NO;

21;

(b) a light chain CDR2 amino acid sequence of SEQ ID NO;

22;

(c) a light chain CDR3 amino acid sequence of LGh;

wherein h is a hydrophobic amino acid;

(d) a heavy chain CDR1 amino acid sequence of YXI, wherein X is any amino acid;

(e) a heavy chain CDR2 amino acid sequence of SEQ ID NO;

23; and

(f) a heavy chain CDR3 amino acid sequence of SEQ ID NO;

24.

2. The isolated antibody of claim 1, wherein the antibody comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

14 and SEQ ID NO;

16.

3. The isolated antibody of claim 1, wherein the antibody comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

13 and SEQ ID NO;

15.

4. The isolated antibody of claim 1, wherein the antibody is encoded by a nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:

17, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

7. The isolated antibody of claim 1, wherein the antibody is selected from the group consisting of a single-chain antibody, an antibody fragment, a chimeric antibody, or a humanized antibody.

8. The isolated antibody of claim 1, wherein the antibody specifically binds to an acetaminophen protein adduct more than 2000 times more effectively than free acetaminophen.

9. The isolated antibody of claim 1, wherein the antibody specifically binds to an acetaminophen protein adduct about 8000 times more effectively than free acetaminophen.

5. An isolated antibody, wherein the antibody specifically binds an acetaminophen-protein adduct but does not specifically bind free acetaminophen and comprises:

(a) a light chain CDR1 amino acid sequence of SEQ ID NO;

21;

(b) a light chain CDR2 amino acid sequence of SEQ ID NO;

22;

(c) a light chain CDR3 amino acid sequence of LGh;

wherein h is a hydrophobic amino acid;

(d) a heavy chain CDR1 amino acid sequence of YXI, wherein X is any amino acid;

(e) a heavy chain CDR2 amino acid sequence of SEQ ID NO;

23; and

(f) a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO;

6 with zero to two amino acid substitutions or SEQ ID NO;

12 with zero to two amino acid substitutions.

10. The isolated antibody of claim 5, wherein the antibody is selected from the group consisting of a single-chain antibody, an antibody fragment, a chimeric antibody, or a humanized antibody.

11. The isolated antibody of claim 5, wherein the antibody specifically binds to an acetaminophen protein adduct more than 2000 times more effectively than free acetaminophen.

12. The isolated antibody of claim 5, wherein the antibody specifically binds to an acetaminophen protein adduct about 8000 times more effectively than free acetaminophen.

6. An isolated antibody, wherein the antibody specifically binds acetaminophen-protein adduct but does not specifically bind free acetaminophen and comprises:

(a) a light chain CDR1 amino acid sequence of SEQ ID NO;

21;

(b) a light chain CDR2 amino acid sequence of SEQ ID NO;

22;

(c) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO;

3 with zero to two amino acid substitutions or SEQ ID NO;

9 with zero to two amino acid substitutions;

(d) a heavy chain CDR1 amino acid sequence of YXI, wherein X is any amino acid;

(e) a heavy chain CDR2 amino acid sequence of SEQ ID NO;

23; and

(f) a heavy chain CDR3 amino acid sequence of SEQ ID NO;

24.

13. The isolated antibody of claim 6, wherein the antibody is selected from the group consisting of a single-chain antibody, an antibody fragment, a chimeric antibody, or a humanized antibody.

14. The isolated antibody of claim 6, wherein the antibody specifically binds to an acetaminophen protein adduct more than 2000 times more effectively than free acetaminophen.

15. The isolated antibody of claim 6, wherein the antibody specifically binds to an acetaminophen protein adduct about 8000 times more effectively than free acetaminophen.