Development of enzyme-linked immunosorbent assays for measurement of leukotrienes and prostaglandins.
Enzyme-linked immunosorbent assays (ELISAs) were developed for the leukotrienes LTC4 and LTB4 and the prostaglandins 6-keto PGF1 alpha and thromboxane (TxB2). In an indirect assay procedure for all 4 eicosanoids a BSA conjugate of the leukotrienes or an ovalbumin conjugate of the prostaglandins was absorbed to polystyrene microtiter plates. Samples containing the respective eicosanoids were incubated in the coated wells with specific rabbit antisera. The wells were then incubated successively with a goat anti-rabbit antibody linked to fluorescein and a rabbit anti-fluorescein antibody linked to alkaline phosphatase. The resultant assays for LTC4, LTB4, 6-keto PGF1 alpha, and TxB2, gave steep, sensitive inhibition curves; IC50s were 0.2, 10, 1, and 0.4 pmol respectively with minimal cross-reactivity to other eicosanoids. The sensitivities and specificities were comparable to those found in the RIA, and the levels determined in this assay correlate well with those determined in non-immunological assays.
Miller
D K
DK
Sadowski
S
S
DeSousa
D
D
Maycock
A L
AL
Lombardo
D L
DL
Young
R N
RN
Hayes
E C
EC
D015121:6-Ketoprostaglandin F1 alpha / Q000032:analysis; D000818:Animals; D001681:Biological Assay; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007975:Leukotriene B4 / Q000032:analysis*; D009504:Neutrophils / Q000187:drug effects; D011453:Prostaglandins / Q000032:analysis*; D051381:Rats; D013189:SRS-A / Q000032:analysis*; D013582:Synovial Fluid / Q000032:analysis; D013929:Thromboxane B2 / Q000032:analysis
D011453:Prostaglandins; D013189:SRS-A; D007975:Leukotriene B4; D013929:Thromboxane B2; D015121:6-Ketoprostaglandin F1 alpha