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US-20260126445-A1 - Assay for Assessing Cancer

US20260126445A1US 20260126445 A1US20260126445 A1US 20260126445A1US-20260126445-A1

Abstract

Described herein are immunoassay methods for detecting and/or monitoring a cancer in a patient. In the method a biofluid sample from a patient is contacted with a monoclonal antibody that specifically binds to a C-terminal epitope of type XXVIII collagen, and the amount of binding between the monoclonal antibody and peptides in the sample is detected and determined.

Inventors

  • Federica Genovese
  • Nicholas Willumsen
  • Alexander Lynge Reese-Petersen
  • Shu Sun
  • Morten Asser Karsdal

Assignees

  • NORDIC BIOSCIENCE A/S

Dates

Publication Date
20260507
Application Date
20250813
Priority Date
20200911

Claims (10)

  1. 1 . A method of immunoassay for detecting and/or monitoring a cancer in a patient, the method comprising: (i) contacting a biofluid sample from a patient with a monoclonal antibody that specifically binds to a C-terminal epitope of type XXVIII collagen, (ii) detecting and determining the amount of binding between said monoclonal antibody and peptides in the sample, and (iii) correlating said amount of binding of said monoclonal antibody as determined in step (ii) with values associated with normal healthy subjects and/or values associated with known cancer severity and/or values obtained from said patient at a previous time point and/or a predetermined cut-off value.
  2. 2 . The method of claim 1 , wherein the cancer is lung, breast, colorectal or pancreatic cancer.
  3. 3 . The method of claim 2 , wherein the method is a method for detecting the stage or determining the prognosis of a cancer in a patient.
  4. 4 . The method of claim 1 , wherein said monoclonal antibody specifically binds to a C-terminus amino acid sequence QETCIQG (SEQ ID NO: 1).
  5. 5 . The method of claim 4 , wherein said monoclonal antibody does not recognize or specifically bind to an elongated version of said C-terminus amino acid sequence which is QETCIQGA (SEQ ID NO: 2).
  6. 6 . The method of claim 4 , wherein said monoclonal antibody does not recognize or specifically bind to a truncated version of said C-terminus amino acid sequence which is QETCIQ (SEQ ID NO: 3).
  7. 7 . The method of claim 4 , wherein the monoclonal antibody is raised against a synthetic peptide having the amino acid sequence QETCIQG (SEQ ID NO: 1).
  8. 8 . The method of claim 1 , wherein said biofluid is blood, serum, plasma, urine or a supernatant from cell or tissue cultures.
  9. 9 . The method of claim 1 , wherein said immunoassay is a competition assay or a sandwich assay.
  10. 10 . The method of claim 1 , wherein said immunoassay is a radioimmunoassay or an enzyme-linked immunosorbent assay.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This continuation-in-part claims benefit of priority under 35 U.S.C. § 120 of pending application U.S. Ser. No. 18/025,704, filed Mar. 10, 2023, which is a national stage application under 35 U.S.C § 371 of international application PCT/EP2021/074996, filed Sep. 10, 2021, now abandoned, which claims benefit of priority under 35 U.S.C. § 119 (a) of European patent application 2014323.6, filed Sep. 11, 2020, now abandoned, the entirety of all of which are hereby incorporated by reference. INCORPORATION BY REFERENCE OF SEQUENCE LISTING The Sequence Listing XML file, entitled D7956CIPSEQ.xml, was created on Aug. 12, 2025, and has a size of 19000 bytes. This Sequence Listing is hereby incorporated by reference in its entirety. BACKGROUND OF THE INVENTION Field of the Invention The present invention relates to immunoassays for detecting and/or monitoring a cancer in a patient. The cancer may in particular be lung, breast, colorectal or pancreatic cancer. The immunoassay may in certain embodiments be for detecting the stage of a cancer in a patient or determining the prognosis of a cancer in a patient. Description of the Related Art Type XXVIII collagen is poorly described in literature but research entailing its physical role is slowly emerging. It is mainly located in peripheral nerves and dorsal root ganglia, but is also found in the skin (1,2). Type XXVIII collagen is a beaded collagen that structurally resembles type VI collagen, with two von Willebrand factor A domains flanking a 528 amino acid collagenous domain (3). Type XXVIII collagen was found in very low levels in healthy lung tissue but was overexpressed in bleomycin-induced lung injury (4), which could indicate that cells expressing type XXVIII collagen might be involved in tissue repair processes. Type XXVIII collagen has also previously been seen to be upregulated in mouse hepatocarcinoma (5). SUMMARY OF THE INVENTION The present inventors have now determined that collagen type XXVIII formation is upregulated in cancers, such as in particular lung, breast, colorectal or pancreatic cancers, and have developed a competitive ELISA utilizing monoclonal antibodies targeting the C-terminal end of type XXVIII collagen that can be used to detect and/or monitor a cancer and determine the stage of and/or prognosis of a cancer. Accordingly, the present invention provides a method of immunoassay for detecting and/or monitoring a cancer in a patient, the method comprising: (i) contacting a biofluid sample from a patient with a monoclonal antibody that specifically binds to a C-terminal epitope of type XXVIII collagen,(ii) detecting and determining the amount of binding between said monoclonal antibody and peptides in the sample or samples, and(iii) correlating said amount of binding of said monoclonal antibody as determined in step (ii) with values associated with normal healthy subjects and/or values associated with known cancer severity and/or values obtained from said patient at a previous time point and/or a predetermined cut-off value. The immunoassay may be, but is not limited to, a competition assay or a sandwich assay. The immunoassay may, for example, be a radioimmunoassay or an enzyme-linked immunosorbent assay (ELISA). Such assays are techniques known to the person skilled in the art. The cancer may in certain embodiments be lung, breast, colorectal, ovarian and/or pancreatic cancer. In particular, the cancer may be lung, breast, colorectal and/or pancreatic cancer. The method may in certain embodiments be a method for detecting the severity of a cancer in a patient. For example, the method may be a method for detecting the stage of a cancer in a patient, and/or a method of determining the prognosis of a cancer in a patient (such as for example determining the likely survival time or probability of survival of the patient). In certain embodiments, the patient may for example be a patient undergoing a therapy for the cancer, and the method may comprise monitoring the cancer in the patient. The patient biofluid sample may be, but is not limited to, blood, serum, plasma, urine or a supernatant from cell or tissue cultures. Preferably the biofluid is serum or plasma, most preferably serum. As used herein the term “monoclonal antibody” refers to both whole antibodies and to fragments thereof that retain the binding specificity of the whole antibody, such as for example a Fab fragment, F(ab′) 2 fragment, single chain Fv fragment, or other such fragments known to those skilled in the art. As is well known, whole antibodies typically have a “Y-shaped” structure of two identical pairs of polypeptide chains, each pair made up of one “light” and one “heavy” chain. The N-terminal regions of each light chain and heavy chain contain the variable region, while the C-terminal portions of each of the heavy and light chains make up the constant region. The variable region comprises three complementarity determining regions (CDRs