Advanced

Novel COMP Neoepitopes Identified in Synovial Fluids from Patients with Joint Diseases using Affinity Chromatography and Mass Spectrometry.

Åhrman, Emma LU ; Lorenzo, Pilar LU ; Holmgren, Kristin LU ; Grodzinsky, Alan LU ; Dahlberg, Leif LU ; Saxne, Tore LU ; Heinegård, Dick LU and Önnerfjord, Patrik LU (2014) In Journal of Biological Chemistry 289(30). p.20908-20916
Abstract
To identify patients at risk for progressive joint damage there is a need for early diagnostic tools to detect molecular events leading to cartilage destruction. Isolation and characterization of distinct cartilage oligomeric matrix protein (COMP) fragments derived from cartilage and released into synovial fluid will allow discrimination between different pathological conditions and monitoring of disease progression. Early detection of disease and processes in the tissue as well as an understanding of the pathologic mechanisms will also open for novel treatment strategies. Disease specific COMP fragments were isolated by affinity chromatography of synovial fluids from patients with rheumatoid arthritis (RA), osteoarthritis (OA) or acute... (More)
To identify patients at risk for progressive joint damage there is a need for early diagnostic tools to detect molecular events leading to cartilage destruction. Isolation and characterization of distinct cartilage oligomeric matrix protein (COMP) fragments derived from cartilage and released into synovial fluid will allow discrimination between different pathological conditions and monitoring of disease progression. Early detection of disease and processes in the tissue as well as an understanding of the pathologic mechanisms will also open for novel treatment strategies. Disease specific COMP fragments were isolated by affinity chromatography of synovial fluids from patients with rheumatoid arthritis (RA), osteoarthritis (OA) or acute trauma (AT). Enriched COMP fragments were separated by SDS-PAGE followed by in-gel digestion and mass spectrometric identification and characterization. Using the enzymes trypsin, chymotrypsin and Asp-N for the digestions an extensive analysis of the enriched fragments could be accomplished. Twelve different neoepitopes were identified and characterized within the enriched COMP fragments. For one of the neoepitopes, S77, an inhibition ELISA was developed. This ELISA quantifies COMP fragments clearly distinguishable from total COMP. Furthermore, fragments containing the neoepitope S77 were released into the culture medium of cytokine (TNF-α and IL-6/sIL-6R) stimulated human cartilage explants. The identified neoepitopes provide a complement to the currently available commercial assays for cartilage markers. Through neoepitope assays, tools to pin-point disease progression, evaluation methods for therapy and means to elucidate disease mechanisms will be provided. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
289
issue
30
pages
20908 - 20916
publisher
ASBMB
external identifiers
  • pmid:24917676
  • wos:000339396600044
  • scopus:84905372439
ISSN
1083-351X
DOI
10.1074/jbc.M114.554683
language
English
LU publication?
yes
id
8582a0c2-5918-4f9c-9356-dbddc090129a (old id 4528815)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/24917676?dopt=Abstract
date added to LUP
2014-07-05 17:23:53
date last changed
2017-11-12 03:07:39
@article{8582a0c2-5918-4f9c-9356-dbddc090129a,
  abstract     = {To identify patients at risk for progressive joint damage there is a need for early diagnostic tools to detect molecular events leading to cartilage destruction. Isolation and characterization of distinct cartilage oligomeric matrix protein (COMP) fragments derived from cartilage and released into synovial fluid will allow discrimination between different pathological conditions and monitoring of disease progression. Early detection of disease and processes in the tissue as well as an understanding of the pathologic mechanisms will also open for novel treatment strategies. Disease specific COMP fragments were isolated by affinity chromatography of synovial fluids from patients with rheumatoid arthritis (RA), osteoarthritis (OA) or acute trauma (AT). Enriched COMP fragments were separated by SDS-PAGE followed by in-gel digestion and mass spectrometric identification and characterization. Using the enzymes trypsin, chymotrypsin and Asp-N for the digestions an extensive analysis of the enriched fragments could be accomplished. Twelve different neoepitopes were identified and characterized within the enriched COMP fragments. For one of the neoepitopes, S77, an inhibition ELISA was developed. This ELISA quantifies COMP fragments clearly distinguishable from total COMP. Furthermore, fragments containing the neoepitope S77 were released into the culture medium of cytokine (TNF-α and IL-6/sIL-6R) stimulated human cartilage explants. The identified neoepitopes provide a complement to the currently available commercial assays for cartilage markers. Through neoepitope assays, tools to pin-point disease progression, evaluation methods for therapy and means to elucidate disease mechanisms will be provided.},
  author       = {Åhrman, Emma and Lorenzo, Pilar and Holmgren, Kristin and Grodzinsky, Alan and Dahlberg, Leif and Saxne, Tore and Heinegård, Dick and Önnerfjord, Patrik},
  issn         = {1083-351X},
  language     = {eng},
  number       = {30},
  pages        = {20908--20916},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Novel COMP Neoepitopes Identified in Synovial Fluids from Patients with Joint Diseases using Affinity Chromatography and Mass Spectrometry.},
  url          = {http://dx.doi.org/10.1074/jbc.M114.554683},
  volume       = {289},
  year         = {2014},
}