Development and Evaluation of a Microtiter Plate Enzyme Immunoassay for Antibodies to 3,3′-Dichlorobenzidine
| Autor: | Vogt R, F W Spierto, Powell Mk, F H Gardner, W H Hannon, W Whitfield |
|---|---|
| Rok vydání: | 1987 |
| Předmět: |
Health
Toxicology and Mutagenesis Radioimmunoassay 3 3'-Dichlorobenzidine Toxicology Horseradish peroxidase Antibodies Analytical Chemistry Immunoenzyme Techniques Microtiter plate chemistry.chemical_compound Antibody Specificity medicine Animals Environmental Chemistry Bovine serum albumin Antiserum Chemical Health and Safety Chromatography biology medicine.diagnostic_test Benzidines Antibody titer chemistry Immunoassay biology.protein Rabbits |
| Zdroj: | Journal of Analytical Toxicology. 11:31-35 |
| ISSN: | 1945-2403 0146-4760 |
| DOI: | 10.1093/jat/11.1.31 |
| Popis: | The authors obtained and evaluated antisera from rabbits injected with a derivative of a potent bladder carcinogen, dichlorobenzidine (DCB), conjugated to bovine serum albumin (BSA). A 14C-radioimmunoassay (RIA) was able to detect the presence of DCB antibodies, but its relative insensitivity led to the development of a more sensitive enzyme immunoassay (EIA). The EIA test was a "sandwich" method in which a second antibody, labeled with an enzyme (horseradish peroxidase), was used to measure antibody binding to transferrin (Tf)-conjugated DCB immobilized on a microtiter plate. Antibody titers measured by RIA were approximately 1:40; when measured by EIA, they were approximately 1:40,000. Antibody specificity was assessed by comparing the antibody binding activities of DCB, BSA, Tf, BSA-conjugated to DCB, and a number of N-substituted aromatic compounds that included benzidine (Bz). Among the compounds tested, the rabbit antiserum reacted only with DCB and the carrier protein, BSA. Moreover, antibody binding activity to Tf-conjugated DCB was significantly inhibited by unconjugated DCB concentrations between 30 and 500 ng/mL. The precision of antibody binding activities as a function of DCB concentration (expressed by the CV) ranged from 9% for low (30 ng/mL) DCB levels to 12% for higher (500 ng/mL) levels. This evaluation suggests that the antiserum obtained would be appropriate for detecting DCB levels at the ng/mL level. |
| Databáze: | OpenAIRE |
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