Immunoaffinity purification of prostaglandin E2 and leukotriene C4 prior to radioimmunoassay: application to human synovial fluid.
When human synovial fluid as such was subjected to radioimmunoassays of prostaglandin E2 (PGE2) and leukotriene C4 (LTC4), there was no linear increase in PGE2 and LTC4 as the amount of synovial fluid was raised. For removal of substances thus disturbing the assay we developed a method of immunoaffinity purification of PGE2 and LTC4. A monoclonal antibody against PGE2 or LTC4 was coupled to BrCN-activated Sepharose 4B. When synovial fluid mixed with radiolabelled PGE2 or LTC4 was applied to the column of immobilized antibody, the ligand was adsorbed to the column and eluted with a mixture of methanol/water in a recovery of about 80%. The purified material showed a linearity between the amount of the sample and the value of radioimmunoassay. The one-step method was applied to synovial fluid from patients with rheumatoid arthritis, osteoarthritis and other joint diseases.
Matsumoto
S
S
Department of Orthopaedic Surgery, Tokushima University, School of Medicine, Japan.
Hayashi
Y
Y
Kinoshita
I
I
Ikata
T
T
Yamamoto
S
S
D000818:Animals; D000911:Antibodies, Monoclonal; D001172:Arthritis, Rheumatoid / Q000175:diagnosis; D002846:Chromatography, Affinity; D015232:Dinoprostone / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D007592:Joint Diseases / Q000175:diagnosis; D051379:Mice; D008807:Mice, Inbred BALB C; D010003:Osteoarthritis / Q000175:diagnosis; D011863:Radioimmunoassay; D013189:SRS-A / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D013582:Synovial Fluid / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D013189:SRS-A; D015232:Dinoprostone