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US-12624121-B2 - Assays for TIMP2 having improved performance in biological samples

US12624121B2US 12624121 B2US12624121 B2US 12624121B2US-12624121-B2

Abstract

The present invention relates to antibodies or antigen binding fragments thereof that binds to human “TIMP2. Further provided are methods for treating subjects, including human subjects, in need of treatment with the isolated TIMP2 antibodies or antigen-binding fragments thereof disclosed herein. Further provided are pharmaceutical or sterile compositions of anti-TIMP2 antibodies and antigen-binding fragments of the invention, the antibody or antigen-binding fragment thereof is admixed with a pharmaceutically acceptable carrier or excipient. Further provided are kits comprising one or more components that include an anti-TIMP2 antibody or antigen-binding fragment thereof of the invention or a pharmaceutical composition thereof.

Inventors

  • Ravi A. VIJAYENDRAN
  • Srivatsa Venkatasubbarao

Assignees

  • BIOMERIEUX, INC.

Dates

Publication Date
20260512
Application Date
20230621

Claims (20)

  1. 1 . A monoclonal antibody that binds to human tissue inhibitor of metalloproteinases 2 (TIMP2) protein, wherein the antibody comprises: (a) a light chain variable region complementarity determining region 1 (CDR1) comprising the amino acid sequence DHINNW (SEQ ID NO: 24), a light chain variable region CDR2 comprising the amino acid sequence SGA, and a light chain variable region CDR3 comprising the amino acid sequence QQYWSTPFT (SEQ ID NO: 26); and (b) a heavy chain variable region CDR1 comprising the amino acid sequence GYSFTSYW (SEQ ID NO: 27), a heavy chain variable region CDR2 comprising the amino acid sequence IDPSDSET (SEQ ID NO: 28), and a heavy chain variable region CDR3 comprising the amino acid sequence ARRDYGSRYDAMDY (SEQ ID NO: 29).
  2. 2 . The monoclonal antibody of claim 1 , wherein the light chain variable region comprises an amino acid sequence at least 90% identical to that set forth as SEQ ID NO: 30 and the heavy chain variable region comprises an amino acid sequence at least 90% identical to that set forth as SEQ ID NO: 31.
  3. 3 . The monoclonal antibody of claim 2 , wherein the light chain variable region comprises an amino acid sequence set forth as SEQ ID NO: 30 and the heavy chain variable region comprises an amino acid sequence set forth as SEQ ID NO: 31.
  4. 4 . The monoclonal antibody of claim 1 , wherein the light chain comprises an amino acid sequence at least 90% identical to that set forth as SEQ ID NO: 39 and the heavy chain comprises an amino acid sequence at least 90% identical to that set forth as SEQ ID NO: 33.
  5. 5 . The monoclonal antibody of claim 4 , wherein the light chain comprises an amino acid sequence set forth as SEQ ID NO: 39 and the heavy chain comprises an amino acid sequence set forth as SEQ ID NO: 33.
  6. 6 . The monoclonal antibody of claim 1 , wherein the antibody is a rabbit antibody.
  7. 7 . The monoclonal antibody of claim 1 , wherein the antibody is conjugated to a signal development element.
  8. 8 . The monoclonal antibody of claim 1 , wherein the antibody is immobilized on a solid support.
  9. 9 . The monoclonal antibody of claim 1 , wherein one of the first and the second monoclonal antibodies is immobilized on a solid support.
  10. 10 . A rabbit monoclonal antibody comprising a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 30 and a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 31.
  11. 11 . A method for determining the presence or amount of human tissue inhibitor of metalloproteinases 2 (TIMP2) protein in a biological sample, the method comprising: performing an immunoassay on the biological sample with first and second monoclonal antibodies which together form sandwich complexes with human TIMP2 protein, wherein the immunoassay provides a detectable signal that is related to the presence or amount of human TIMP2 protein in the biological sample bound in the sandwich complexes; and relating the detectable signal to the presence or amount of human TIMP2 protein in the biological sample, wherein the first monoclonal antibody comprises: (a) a light chain variable region complementarity determining region 1 (CDR1) comprising the amino acid sequence DHINNW (SEQ ID NO: 24), a light chain variable region CDR2 comprising the amino acid sequence SGA (SEQ ID NO: 25), and a light chain variable region CDR3 comprising the amino acid sequence QQYWSTPFT (SEQ ID NO: 26); and (b) a heavy chain variable region CDR1 comprising the amino acid sequence GYSFTSYW (SEQ ID NO: 27), a heavy chain variable region CDR2 comprising the amino acid sequence IDPSDSET (SEQ ID NO: 28), and a heavy chain variable region CDR3 comprising the amino acid sequence ARRDYGSRYDAMDY (SEQ ID NO: 29).
  12. 12 . The method of claim 11 , wherein the first monoclonal antibody comprises a light chain variable region comprising an amino acid sequence at least 90% identical to that set forth as SEQ ID NO: 30 and a heavy chain variable region comprising an amino acid sequence at least 90% identical to that set forth as SEQ ID NO: 31.
  13. 13 . The method of claim 12 , wherein the light chain variable region comprises an amino acid sequence set forth as SEQ ID NO: 30 and the heavy chain variable region comprises an amino acid sequence set forth as SEQ ID NO: 31.
  14. 14 . The method of claim 11 , wherein the first monoclonal antibody comprises a light chain comprising an amino acid sequence at least 90% identical to that set forth as SEQ ID NO: 39 and a heavy chain comprising an amino acid sequence at least 90% identical to that set forth as SEQ ID NO: 33.
  15. 15 . The method of claim 14 , wherein the light chain comprises an amino acid sequence set forth as SEQ ID NO: 39 and the heavy chain comprises an amino acid sequence set forth as SEQ ID NO: 33.
  16. 16 . The method of claim 11 , wherein the first monoclonal antibody is a rabbit antibody.
  17. 17 . The method of claim 11 , wherein one of the first and the second monoclonal antibodies is conjugated to a signal development element.
  18. 18 . The method of claim 11 , wherein the minimum detectable concentration is 10 ng/ml or less.
  19. 19 . The method of claim 11 , wherein the immunoassay is performed in a lateral flow format.
  20. 20 . The method of claim 11 , wherein the immunoassay is performed by applying the biological sample to a disposable test device, and the detectable signal is obtained by inserting the disposable test device into an analytical instrument, wherein the sandwich complexes comprising the first and second antibodies are immobilized for detection in a predetermined zone of the disposable test device, and wherein the analytical instrument detects the immobilized sandwich complexes to provide the detectable signal.

Description

The present application is a divisional application of U.S. patent application Ser. No. 16/603,071 filed Oct. 4, 2019, which is a U.S. National Phase Entry of International Patent Application No. PCT/US2018/026058 filed Apr. 4, 2018, which claims the benefit of U.S. Provisional Patent Application 62/482,089 filed Apr. 5, 2017, which is hereby incorporated by reference in its entirety, including all tables, figures, and claims. SEQUENCE LISTING This application contains a Sequence Listing which has been submitted in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on Jun. 21, 2023, is named AST M_0005_DV_Sequence_Listing.xml and is 66,735 bytes in size. BACKGROUND The following discussion of the background of the invention is merely provided to aid the reader in understanding the invention and is not admitted to describe or constitute prior art to the present invention. Metalloproteinase inhibitor 2 (Swiss-Prot P16035, also known as “Tissue inhibitor of metalloproteinases 2” and “TIMP2”) is a secreted protein which complexes with metalloproteinases and irreversibly inactivates them by binding to their catalytic zinc cofactor. TIMP2 is known to act on MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-13, MMP-14, MMP-15, MMP-16 and MMP-19. TIMP2 reportedly suppresses the proliferation of endothelial cells. As a result, the encoded protein has been suggested to have a role in the maintenance of tissue homeostasis by suppressing the proliferation of quiescent tissues in response to angiogenic factors, and by inhibiting protease activity in tissues undergoing remodeling. In addition, WO2010/048346 and WO2011/075744, each of which is hereby incorporated by reference in its entirety including all tables, figures and claims, describe the use of TIMP2 for evaluating the renal status of a subject both individually and in multimarker panels. In particular, TIMP2 levels measured by immunoassay are shown to correlate to risk stratification, diagnosis, staging, prognosis, classifying and monitoring of the renal status. PCT/US2014/050195 discloses certain antibodies for use in such assays. Signals obtained from specific binding assays such as immunoassays are a direct result of complexes formed between one or more binding species (e.g., antibodies) and the target biomolecule (i.e., the analyte) and polypeptides containing the necessary epitope(s) to which the antibodies bind. Immunoassays are often able to “detect” an analyte; but because an antibody epitope is on the order of 8 amino acids, an immunoassay configured to detect a marker of interest will also detect polypeptides related to the marker sequence, so long as those polypeptides contain the epitope(s) necessary to bind to the antibody or antibodies used in the assay. While such assays may detect the full length biomarker and the assay result be expressed as a concentration of a biomarker of interest, the signal from the assay is actually a result of all such “immunoreactive” polypeptides present in the sample. Such binding assays may also detect immunoreactive polypeptides present in a biological sample that are complexed to additional species, such as binding proteins, receptors, heparin, lipids, sugars, etc., provided that those additional species do not interfere in binding between the binding species and the target biomolecule. Typically, however, specific binding assays are formulated using purified analyte, and complex formation and fragmentation patterns are not considered. This is particularly true where the identity of such additional binding species are unknown. SUMMARY It is an object of the invention to provide antibodies which bind novel epitopes of TIMP2. Such antibodies can find use in immunoassays with improved clinical performance, particularly when used in the evaluation of renal injuries, and in therapeutic methods in which TIMP2 binding is desired. In a first aspect, the present invention relates to antibodies or antigen binding fragments thereof that binds to human TIMP2, wherein the antibody or antigen binding fragment comprises one or more, and optionally each, of: a light chain variable region CDR1 comprising the amino acid sequence ESISGW (SEQ ID NO:1) or an amino acid sequence at least 90%, 95%, 97%, 98%, or 99% identical thereto,a light chain variable region CDR2 comprising the amino acid sequence RAS (SEQ ID NO:2) or an amino acid sequence at least 90%, 95%, 97%, 98%, or 99% identical thereto,a light chain variable region CDR3 comprising the amino acid sequence QCSYGINGNSEHGNP (SEQ ID NO:3) or an amino acid sequence at least 90%, 95%, 97%, 98%, or 99% identical thereto,a heavy chain variable region CDR1 comprising the amino acid sequence GFTISSNYY (SEQ ID NO:4) or an amino acid sequence at least 90%, 95%, 97%, 98%, or 99% identical thereto,a heavy chain variable region CDR2 comprising the amino acid sequence ILGGSGTYT (SEQ ID NO:5) or an amino acid sequence at least 90%, 95%, 97%, 98%, or 99