Date of Award

Summer 1990

Document Type


Degree Name

Master of Science (MS)


Chemistry & Biochemistry



Committee Director

James H. Yuan

Committee Member

Mark S. Elliot

Call Number for Print

Special Collections LD4331.C45Y34


A sensitive and simple microwell based competition enzyme immunoassay for the quantitative determination of estradiol (E2) and estriol (E3) was developed. Nicrowells coated with antibody were incubated with antigen followed by adding horseradish peroxidase (HRPO) conjugate. The assay, which can be performed within two hours at room temperature, involved simultaneous incubation of E2 - or E3-HRPO conjugate and serum sample in polystyrene microwells coated with anti- E2 or anti-E3 gamma globulin fraction.

Gamma globulin was isolated from whole anti-serum by DEAE-cellulose chromatography. A carbodiimide coupling method was utilized to prepare the E2 - and E3-HRPO conjugates.

The detection limit of the E2 assay is 2.0 pg/mL and 6.0 ng/mL for the E3 assay. Precision studies involving pooled serum samples with three different levels of E2 and E3 were performed . Intra-assay coefficients of variation of 15.0%, 8.83%, and 8.13% were obtained for E2 at 20.48 pg/mL, 79.14 pg/mL, and 424.6 pg/mL, and of 12.4%, 9.63%, and 9.08% for E3 at 49.19 ng/mL, 153.4 ng/mL, and 505.7 ng/mL. Interassay coefficients of variation of 18.04&, 10.404, and 8.70% were obtained for E2 at 20.57 pg/mL, 78.67 pg/mL, and 422.0 pg/mL, and of 15.06%, 12.33%, and 9.84% for E, at 47.76 ng/mL, 153.4 ng/mL, and 504.7 ng/mL. Values for total E2 and total E3 so determined correlated well with those determined by radioimmunoassay with correlation coefficient of 0.955 and 0.959 respectively.


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