The present invention provides an immunological binding partner reactive with a C-terminal epitope of the C5 domain of the α3 chain of collagen Type 6, and a method of immunoassay using the immunological binding partner for detecting and quantifying the C-terminal epitope. The invention also provides a method of investigating the rate of formation of extracellular matrix and a method for identifying a subject suitable for treatment with an insulin sensitizer.

An immune checkpoint inhibitor containing a substance that inhibits binding between fibronectin and an immunosuppressive receptor LILRB4 as an active ingredient, a therapeutic agent for an immune checkpoint-related disease, an immunosuppressant containing a substance that activates an immunosuppressive receptor LILRB4 as an active ingredient, an anti-fibronectin antibody or a derivative thereof, a fibronectin analog, a kit for detecting fibronectin or a partial protein thereof, and a method for detecting fibronectin or a partial protein thereof in a biological sample using the kit.

The present disclosure provides methods for isolating tumor-derived microparticles from a subject for analysis, specifically by isolating Tenascin-C positive microparticles from a sample from the subject to obtain tumor-derived microparticles. Methods for determining the expression status of biomarkers in the tumor-derived microparticles and methods for determining additional characteristics of the tumor-derived microparticles are also provided.

A method of immunoassay for detecting and/or monitoring a cardiovascular disease in a patient and/or assessing the likelihood of or the severity of a cardiovascular disease in a patient, comprising contacting a biofluid sample from a patient with a monoclonal antibody that specifically binds to a C-terminal epitope of the C5 domain of the α3 chain of type VI collagen, and/or contacting a biofluid sample from the patient with a monoclonal antibody that specifically binds to a C-terminal neo-epitope of the N-terminal propeptide of type III collagen.

Provided herein are methods for treating a patient having NASH who has been determined to have a particular threshold level of serum Pro-C3 (e.g., greater than 10 ng/ML) by administering to the patient a modified Fibroblast growth factor 21 (FGF-21) in an amount and with a frequency sufficient to treat NASH. Also provided are methods for monitoring responsiveness of a patient having NASH to treatment with a modified FGF-21, the method comprising: determining the serum Pro-C3 level in a blood sample from the patient obtained during or after treatment, wherein: a decreased serum Pro-C3 level in the blood sample from the patient obtained during or after treatment, as compared to the serum Pro-C3 level in a blood sample from the patient obtained prior to treatment with the modified FGF-21, indicates that the patient is responsive to treatment with the modified FGF-21.

An object of the present invention is to provide a novel method that enables determination or evaluation of the undifferentiated state of human pluripotent stem cells simply, efficiently and non-invasively. The present invention provides a method for determining or evaluating the undifferentiated state of human pluripotent stem cells, comprising a step of detecting or measuring fibronectin in a culture supernatant of human pluripotent stem cells.

A method for performing an enzyme-linked immunosorbent assay (ELISA), the method comprising immobilizing a capture antibody to a substrate, wherein the capture antibody is configured to bind to a laminin beta-1 chain; adding a micronized tissue sample to the substrate containing the capture antibody; adding a detection antibody to the substrate containing the capture antibody and the micronized tissue sample, wherein the detection antibody is configured to bind to a laminin gamma-1 chain; detecting whether a complex of the capture antibody, a target antigen in the micronized tissue sample, and the detection antibody has been formed, wherein presence of the complex indicates presence of the target antigen with intact tertiary structure in the micronized tissue sample, and absence of the complex indicates absence of the target antigen with intact tertiary structure in the micronized tissue sample.

Disclosed are peptide conjugates comprising an active agent, a spacer moiety, and a dimeric collagen hybridizing peptide comprising a first and second collagen hybridizing peptide, a linker; and a branch point, wherein the first and second collagen hybridizing peptides comprise the sequence of at least (GXY)n, wherein G is glycine, wherein X and Y are any amino acid, and wherein n is any number between 3 and 12. Also disclosed are methods of detecting denatured collagen in a sample comprising contacting a composition comprising any one of the disclosed peptide conjugates to a sample, wherein the active agent comprises a therapeutic agent, and detecting the presence or absence of binding of the peptide conjugate to denatured collagen, the presence of binding indicating the presence of denatured collagen in the sample. Also disclosed are methods of treating a disease or injury involving collagen damage comprising administering to a subject having a disease or injury involving collagen damage any one of the disclosed peptide conjugates.

Fibronectin type III domains (FN3) that specifically bind to serum albumin, related polynucleotides capable of encoding serum albumin-specific FN3 domains, cells expressing the FN3 domains, as well as associated vectors, detectably labeled FN3 domains and FN3 domains fused to a heterologous moiety are useful in extending the half-life of molecules in diagnostic and therapeutic applications.

Provided herein is a sandwich immunoassay for detecting cross-linked PIIINP that has at least two strands of PIIINP joined together by inter-strand cross-linking each having a C-terminal neo-epitope of PIIINP that is generated by N-protease cleavage of intact type III procollagen. A biological sample having the cross-linked PIIINP is contacted with a first surface-bound monoclonal antibody and then by a second monoclonal antibody, both specifically reactive with a neoepitope in the C-terminal sequence of PIIINP, and then binding of the second monoclonal antibody is determined. Also provided is a method for evaluating the efficacy of an antagonist drug targeting lysyl oxidases via the immunoassay and a kit containing a solid support binding the first monoclonal antibody and containing the second monoclonal antibody.