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Home / Equine Research Bank / Int J Mol Sci / Development of a Cartilage Oligomeric Matrix Protein Neo-Epi...
International journal of molecular sciences2020; 21(6); 2155; doi: 10.3390/ijms21062155

Development of a Cartilage Oligomeric Matrix Protein Neo-Epitope Assay for the Detection of Intra-Thecal Tendon Disease.

Abstract: The diagnosis of tendon injury relies on clinical signs and diagnostic imaging but imaging is subjective and does not always correlate with clinical signs. A molecular marker would potentially offer a sensitive and specific diagnostic tool that could also provide objective assessment of healing for the comparison of different treatments. Cartilage Oligomeric Matrix Protein (COMP) has been used as a molecular marker for osteoarthritis in humans and horses but assays for the protein in tendon sheath synovial fluids have shown overlap between horses affected by tendinopathy and controls. We hypothesized that quantifying a COMP neoepitope would be more discriminatory of injury. COMP fragments were purified from synovial fluids of horses with intra-thecal tendon injuries and media from equine tendon explants, and mass spectrometry of a consistent and abundant fragment revealed a ~100 kDa COMP fragment with a new N-terminus at the 78th amino-acid (NH-TPRVSVRP) located just outside the junctional region of the protein. A competitive inhibition ELISA based on a polyclonal antibody raised to this sequence yielded more than a 10-fold rise in the mean neoepitope levels for tendinopathy cases compared to controls (5.3 ± 1.3 µg/mL ( = 7) versus 58.8 ± 64.3 µg/mL ( = 13); = 0.002). However, there was some cross-reactivity of the neoepitope polyclonal antiserum with intact COMP, which could be blocked by a peptide spanning the neoepitope. The modified assay demonstrated a lower concentration but a significant > 500-fold average rise with tendon injury (2.5 ± 2.2 ng/mL ( = 6) versus 1029.8 ± 2188.8 ng/ml ( = 14); = 0.013). This neo-epitope assay therefore offers a potentially useful marker for clinical use.
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Publication Date: 2020-03-20 PubMed ID: 32245107PubMed Central: PMC7139564DOI: 10.3390/ijms21062155Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research focuses on developing a method that uses Cartilage Oligomeric Matrix Protein (COMP) as a molecular marker to detect tendon injury in horses. The study suggests that this new technique could be far more specific and sensitive in diagnosing such injuries, comparing favourably to current imaging-based methods.

Objective of the Study

  • The objective of the study was to develop a sensitive and specific diagnostic tool for the detection of tendon injury in horses that could possibly replace subjective imaging methods.
  • The researchers chose to test COMP as a potential molecular marker, as previous studies have shown it to be effective in diagnosing osteoarthritis in humans and horses.

Research Methods

  • The researchers hypothesized that quantifying a COMP neoepitope (a new antigen that is generated after a protein undergoes a certain process) would be more indicative of tendon injury.
  • COMP fragments were extracted from the synovial fluid (lubricating fluid between joints) of horses that had tendon injuries (intra-thecal), and from tendon explants in an equine laboratory.
  • A competitive inhibition ELISA (a type of analysis used for measuring the concentration of antibodies in samples) was used to discover the concentration of the new COMP neoepitope.

Results and Findings

  • Through mass spectrometry (a powerful technique used for identifying and quantifying substances within a sample), they found a COMP fragment with a new end at the 78th amino acid.
  • An ELISA based on an antibody raised to this sequence resulted in a more than ten-fold rise in the levels of the neoepitope in the cases of tendinopathy (a tendon disorder) when compared to the control group.
  • There occurred some cross-reactivity between the neoepitope and the intact COMP, this was counteracted with a peptide overlapping the neoepitope.
  • After refinement in the assay, it still showed a significant average rise in concentration (of over 500-fold) during a tendon injury.

Conclusions and Outlook

  • Overall, the study demonstrated that this modified neo-epitope assay could act as a powerful indicator of tendon injury, presenting a potential new diagnostic tool.
  • This research could have considerable impact on clinical methods for treating and managing tendon diseases in horses.

Cite This Article

APA
Smith R, Önnerfjord P, Holmgren K, di Grado S, Dudhia J. (2020). Development of a Cartilage Oligomeric Matrix Protein Neo-Epitope Assay for the Detection of Intra-Thecal Tendon Disease. Int J Mol Sci, 21(6), 2155. https://doi.org/10.3390/ijms21062155

Publication

International journal of molecular sciences
ISSN: 1422-0067
NlmUniqueID: 101092791
Country: Switzerland
Language: English
Volume: 21
Issue: 6
PII: 2155

Researcher Affiliations

Smith, Roger
  • Dept of Clinical Sciences and Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Herts AL9 7TA, UK.
Önnerfjord, Patrik
  • Department of Clinical Sciences, Rheumatology and Molecular Skeletal Biology, University of Lund, 22184 Lund, Sweden.
Holmgren, Kristin
  • Department of Clinical Sciences, Rheumatology and Molecular Skeletal Biology, University of Lund, 22184 Lund, Sweden.
di Grado, Shacko
  • Dept of Clinical Sciences and Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Herts AL9 7TA, UK.
  • Anicura Regiondjursjukhuset Bagarmossen, Ljusnevägen 17, 128 48 Bagarmossen, Sweden.
Dudhia, Jayesh
  • Dept of Clinical Sciences and Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Herts AL9 7TA, UK.

MeSH Terms

  • Amino Acid Sequence
  • Animals
  • Antibodies / immunology
  • Biological Assay / methods
  • Cartilage Oligomeric Matrix Protein / chemistry
  • Cartilage Oligomeric Matrix Protein / immunology
  • Cartilage Oligomeric Matrix Protein / metabolism
  • Cross Reactions / immunology
  • Epitopes / metabolism
  • Horses
  • Spinal Cord / pathology
  • Synovial Fluid / metabolism
  • Tendon Injuries / diagnosis
  • Tendon Injuries / metabolism
  • Tendons / pathology

Grant Funding

  • VET/PRJ/674 and VET/PRJ/703 / Horserace Betting Levy Board

Conflict of Interest Statement

There are no conflicts of interest.

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Citations

This article has been cited 1 times.
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