Enzyme immunosorbant assay of oestradiol in unextracted plasma using penicillinase as label.
An enzyme-linked immunosorbant assay (ELISA) for measuring oestradiol directly in plasma without extraction utilizing antibodies raised against oestradiol-3-(O-carboxymethyl) ether-bovine serum albumin conjugate, and oestradiol-6-(O-carboxymethyl) oxime linked to penicillinase (EC 3.5.2.6) as a marker was developed. Polyvinyl 96-well microtitre plates were used for immobilization of anti-oestradiol IgG. Standards of oestradiol (92 to 9,190 pmol/l were prepared in oestradiol-free plasma and 8-anilino-1-naphthalene sulphonic acid (8-ANS, 5 mg/ml of 10 mmol/l PBS) was added to the microtitre plate wells to displace oestradiol from plasma binding proteins. The assay had a lower limit of detection of 92 pmol/l plasma and could be performed within 4 h. Comparison of oestradiol values of 51 plasma specimens obtained by ELISA with those of radioimmunoassay (RIA), in which oestradiol was extracted with diethyl ether, showed good correlation (y = 0.786x + 0.03; r = 0.900).
Pandey
P K
PK
Department of Reproductive Biomedicine, National Institute of Health and Family Welfare, New Delhi, India.
Shrivastav
T G
TG
Kumari
G L
GL
Rao
P N
PN
Grover
P K
PK
Murthy
H G
HG
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D004958:Estradiol / Q000097:blood*; D005260:Female; D006801:Humans; D008297:Male; D010405:Penicillinase; D011863:Radioimmunoassay; D012016:Reference Values
D004958:Estradiol; D010405:Penicillinase