Development of a radioimmunoassay for prostaglandin D2 using an antiserum against 11-methoxime prostaglandin D2.
A sensitive and specific radioimmunoassay for prostaglandin D2 has been developed using its stabilized 11-methoxime derivative, which was obtained after treatment of prostaglandin D2 with methoxamine-HCl. The antiserum was obtained after injection of prostaglandin D2-methoxamine coupled to bovine serum albumin. A (125I)-Histamide prostaglandin D2-methoxamine tracer was prepared by iodination of the corresponding histamide, followed by thin layer chromatography purification. The sensitivity of the assay was 280 femtomoles per ml at 50% displacement. The cross reactivities were 15% with prostaglandin D1-methoxamine and less than 0.20% with other prostaglandins. Determination of the half-life of prostaglandin D2 in a solution containing albumin was also carried out, since it has been shown to catalyze prostaglandin D2 destruction. The unstability of this prostaglandin is due to the presence of a beta-hydroxy ketone group, and all prostaglandins possessing this labile moiety could be stabilized by such a derivatization before developing a radioimmunoassay.
Maclouf
J
J
Corvazier
E
E
Wang
Z Y
ZY
D002855:Chromatography, Thin Layer; D006801:Humans; D007106:Immune Sera; D007202:Indicators and Reagents; D008832:Microchemistry; D015230:Prostaglandin D2 / Q000031:analogs & derivatives; D011457:Prostaglandins D / Q000097:blood* / Q000276:immunology*; D011465:Prostaglandins, Synthetic / Q000276:immunology*; D011863:Radioimmunoassay / Q000379:methods; D012709:Serum Albumin
D007106:Immune Sera; D007202:Indicators and Reagents; D011457:Prostaglandins D; D011465:Prostaglandins, Synthetic; D012709:Serum Albumin; C048156:11-methoxime prostaglandin D2; D015230:Prostaglandin D2