Der Radioimmunoassay fur Steroidhormone ?Grundlagen, Entwicklung und Anwendung radioimmunologischer Testverfahren

Kyrein, Hans Joachim

doi:10.1007/bf01409671

Cited by 6 publications

(1 citation statement)

References 29 publications

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“…6 To date, the screening assay for 17b-E 2 in bovine plasma or serum, employed as an official technique, is radioimmunoassay (RIA). [7][8][9][10] This method allows rapid, sensitive and inexpensive screening of a large number of samples. However, the major disadvantage of this conventional technique is that it requires radioisotopes and scintillation fluids.…”

Section: Introductionmentioning

confidence: 99%

Development of an electrochemical ELISA for the screening of 17β-estradiol and application to bovine serum

Draisci

Volpe

Compagnone

et al. 2000

Analyst

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A sensitive electrochemical enzyme-linked immunosorbent assay (ELISA) for the detection of 17 beta-estradiol (17 beta-E2) was developed. Optimisation of two ELISA competition assays, using monoclonal or polyclonal antibodies anti-17 beta-estradiol, coupled with the electrochemical detection was firstly performed. The activity of the label enzyme (horseradish peroxidase) was measured electrochemically using 3,3',5,5'-tetramethylbenzidine as substrate. The use of the polyclonal antibody resulted in a more sensitive assay and the detection limit of the assay was estimated to be 20 pg ml-1. The analytical performances of the method were compared to those obtained using a dissociation enhanced lanthanide fluorescence immunoassay (DELFIA). Although sample extraction is not usually required by DELFIA, both extracted and non extracted samples were assayed. The comparison between the two screening techniques revealed similar results for the extracted samples and showed a comparable precision (RSD%), ranging from 6.2 to 13.4 and from 6.7 to 14.3 for DELFIA and ELISA, respectively. The results obtained by these screening assays were confirmed by liquid chromatography atmospheric pressure chemical ionisation tandem mass spectrometry which is currently used to confirm illegal hormone administration for regulatory purposes. The electrochemical enzyme immunoassay appears suitable as a screening tool for routine analysis of bovine serum estradiol and can be extended to other anabolic hormones using appropriate antibodies.

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Section: Introductionmentioning

confidence: 99%

Development of an electrochemical ELISA for the screening of 17β-estradiol and application to bovine serum

Draisci

Volpe

Compagnone

et al. 2000

Analyst

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Development of novel IC-ELISA as a primary high throughput screening for various estrogen molecules

Cha

Sohn

Jung

et al. 2010

Toxicol. Environ. Health. Sci.

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An indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) has been developed for detecting estrogen molecules in environmental samples.For generating anti-estrogen monoclonal antibody (mAb), BALB/c mice were immunized with 17β-estradiol (E2)-BSA and 5760 hybridoma cell lines were established. Through the optimization processes, a mAb (4BSA-e 3C11) and estriol (E3)-BSA were finally selected as a primary antibody and a coating antigen, respectively. The IC 50 value for a standard estrogen (17β-E2) was 6.26 ng mL 1 and the detection range (20-80% B/B 0 ) was 0.01-377.92 ng mL 1 . The developed IC-ELISA showed some cross-reactivities (CRs) to various estrogen analogues, such as estrone (E1) (1.79%), E3 (77.34%), 16-epiestriol (27.54%) and 16 keto-17β-E2 (2.02%). On the other hand, the assay showed a negligible CRs to other steroid hormones (CRs⁄0.063%), suggesting the specificity of the assay to estrogen molecules. For assay validation, the developed IC-ELISA was compared side by side with high performance liquid chromatography (HPLC), which showed no significant difference in their performances between the two methods. The sensitivity of our IC-ELISA was approximately 100 fold higher than that of HPLC. The estrogen contents (Estrogen Equivalent Concentrations; EEC) in field samples were determined using the IC-ELISA, including swine sewage effluents (7.043 0.023 ng-EEC mL 1 ), bovine feces (0.013 0.001 ng-EEC mL 1 ), and avian feces (0.017 0.001 ng-EEC mL 1 ). Conclusively, we have developed an IC-ELISA that is highly sensitive to estrogens as well as can detect various estrogen analogues at the same time. This assay can be used as a primary screening for a large number of field samples before the instrumental analysis that is laborintensive and time-consuming.

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Test strip enzyme immunoassays and the fast screening of nortestosterone and clenbuterol residues in urine samples at the parts per billion level

Ploum¹,

Haasnoot²,

Paulussen³

et al. 1991

Journal of Chromatography B: Biomedical Sciences and Applicatio

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Der Radioimmunoassay fur Steroidhormone ?Grundlagen, Entwicklung und Anwendung radioimmunologischer Testverfahren

Cited by 6 publications

References 29 publications

Development of an electrochemical ELISA for the screening of 17β-estradiol and application to bovine serum

Development of an electrochemical ELISA for the screening of 17β-estradiol and application to bovine serum

Development of novel IC-ELISA as a primary high throughput screening for various estrogen molecules

Test strip enzyme immunoassays and the fast screening of nortestosterone and clenbuterol residues in urine samples at the parts per billion level

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