Sensitive ELISA for determination of serum E2 using a new tracer E2-Biotin.
A new tracer conjugate of E2-Biotin, with different spacers, was synthesized at position 3 in the estradiol molecule for first time. Immunoreactivity of the tracer was determined by reacting with the anti-E2 monoclonal antibody. The monoclonal antibodies raised against E2 were characterized for its use in ELISA detection systems of serum E2. The purified antibody has a high affinity and specificity for E2. The antibody and tracer were used for establishing a competitive ELISA for estradiol (E2). The experimental results showed that the dose-response curve of the assay covered a range of 33-20,000 pg/mL (n = 8). The detection limit is 28.3 pg/mL (S/N = 3). The intra- and inter-assay coefficients of variation for the assay of serum samples ranged from 5.7 to 13.2% and from 5.3 to 10.6%, respectively. Precoated microtiter plates were dried at 4 degrees C and they were stable for up to 3 months.
Zhao
Jinfu
J
The Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing, PR China.
Wang
Yongcheng
Y
Mi
Jianqiu
J
Li
Yuanzong
Y
Chang
Wenbao
W
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001360:Avidin; D001710:Biotin / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004958:Estradiol / Q000031:analogs & derivatives* / Q000097:blood* / Q000276:immunology; D006735:Horseradish Peroxidase; D006801:Humans; D007201:Indicator Dilution Techniques; D015394:Molecular Structure; D015203:Reproducibility of Results; D013997:Time Factors
D000911:Antibodies, Monoclonal; D001360:Avidin; D004958:Estradiol; D001710:Biotin; D006735:Horseradish Peroxidase