Two-dimensional gel electrophoresis of synovial fluid: Method for detecting candidate protein markers for osteoarthritis

Hiroshi Yamagiwa, Gobinda Sarkar, M. Cristine Charlesworth, Daniel J. McCormick, Mark E. Bolander

Research output: Contribution to journalArticle

46 Scopus citations

Abstract

Synovial fluid (SF) is a dynamic reservoir for proteins originating from serum, synovial tissue, and cartilage. The composition of the SF proteome may reflect the pathophysiological conditions affecting the circulatory system and cartilage. Our long-term goal is to identify reliable protein markers for osteoarthritis (OA) in SF. We first evaluated the pattern of SF proteins on two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) as a function of protein loading, pH range for isoelectric focusing, and concentration of acrylamide in SDS-PAGE. Removal of albumin and γ-globulins from the samples did not improve the detection of protein spots on 2D-PAGE. The repeatability of protein spot intensity was tested by triplicate 2D-PAGE of a given sample; these experiments showed low intrasample variability (correlation coefficients 0.89-0.95). Differences between multiple samples were tested by comparing the 2D-PAGE of four samples. These experiments showed slightly greater variation between samples (correlation coefficients 0.85-0.93) and a number of differentially expressed proteins. The intensity of 18 protein spots differed more than fivefold, and the intensity of nine protein spots differed more than 100-fold. These results show that 2D-PAGE can be used under standard conditions to screen SF samples and identify a small subset of proteins in SF that are potential markers associated with OA.

Original languageEnglish (US)
Pages (from-to)482-490
Number of pages9
JournalJournal of Orthopaedic Science
Volume8
Issue number4
DOIs
StatePublished - Jan 1 2003

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Keywords

  • 2D-PAGE
  • Biochemical markers
  • Osteoarthritis
  • Synovial fluid

ASJC Scopus subject areas

  • Surgery
  • Orthopedics and Sports Medicine

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