[Establishment of the dot immunoenzyme filtration assay for quantitative detection of estriol].
OBJECTIVE
To make anti-estriol (E3) monoclonal antibody (mAb) with high specificity, and develop a rapid, simple and sensitive method for the detection of E3 in county-level institutions.


METHODS
The compounds of E3 were conjugated to horseradish peroxidase (HRP), bonine serum albumin (BSA) and ovalbumin (OVA) respectively by the method of active ester, and the hybridoma cell lines secreting specific anti-E3 mAb were developed via monoclonal antidody technology. The dot immunoenzyme filtration assay in competitive-inhibition format was established with nitrocellulose membrane as solid-phase carrier and anti-E3 mAb as coating antibody.


RESULTS
5 hybridoma cell lines secreting specific anti-E3 mAb were obtained with E3-mAb titers in the range of 1×10(5); to 5×10(5); and the affinity constant (Ka) from 5.1×10(8); L/mol to 5.2×10(9); L/mol. The limit of detecting (LOD) value for E3 was 2 ng/mL and the detection range was 2-200 ng/mL.


CONCLUSIONS
The dot immunoenzyme filtration assay with high-speed detection, easy accessibility and high sensitivity was successfully established.
College of Life Science, Northwest University, Xi'an, China.
Tian
T
Xue
Yijun
Y
Zhao
Jie
J
Dong
Hongmin
H
Li
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000918:Antibody Specificity; D002460:Cell Line; D004964:Estriol / Q000032:analysis* / Q000276:immunology; D005260:Female; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D004964:Estriol