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 language | English (US) |
|---|---|
| Pages (from-to) | 482-490 |
| Number of pages | 9 |
| Journal | Journal of Orthopaedic Science |
| Volume | 8 |
| Issue number | 4 |
| DOIs | |
| State | Published - 2003 |
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Keywords
- 2D-PAGE
- Biochemical markers
- Osteoarthritis
- Synovial fluid
ASJC Scopus subject areas
- Surgery
Cite this
Two-dimensional gel electrophoresis of synovial fluid : Method for detecting candidate protein markers for osteoarthritis. / Yamagiwa, Hiroshi; Sarkar, Gobinda; Charlesworth, M. Cristine; Mc Cormick, Daniel J; Bolander, Mark E.
In: Journal of Orthopaedic Science, Vol. 8, No. 4, 2003, p. 482-490.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Two-dimensional gel electrophoresis of synovial fluid
T2 - Method for detecting candidate protein markers for osteoarthritis
AU - Yamagiwa, Hiroshi
AU - Sarkar, Gobinda
AU - Charlesworth, M. Cristine
AU - Mc Cormick, Daniel J
AU - Bolander, Mark E.
PY - 2003
Y1 - 2003
N2 - 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.
AB - 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.
KW - 2D-PAGE
KW - Biochemical markers
KW - Osteoarthritis
KW - Synovial fluid
UR - http://www.scopus.com/inward/record.url?scp=0042513861&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0042513861&partnerID=8YFLogxK
U2 - 10.1007/s00776-003-0657-3
DO - 10.1007/s00776-003-0657-3
M3 - Article
C2 - 12898298
AN - SCOPUS:0042513861
VL - 8
SP - 482
EP - 490
JO - Journal of Orthopaedic Science
JF - Journal of Orthopaedic Science
SN - 0949-2658
IS - 4
ER -