Development of ultra-sensitive soybean peroxidase-based CL-ELISA for the determination of human thyroglobulin
| Autor: | Marina M. Vdovenko, Alexander V. Zubkov, Leopoldo Della Ciana, Nina S. Kuzmina, Galina I. Kuznetsova, Ivan Yu. Sakharov |
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| Rok vydání: | 2010 |
| Předmět: |
medicine.medical_specialty
medicine.medical_treatment Immunology Enzyme-Linked Immunosorbent Assay Sensitivity and Specificity Thyroglobulin Horseradish peroxidase law.invention law Internal medicine medicine Humans Immunology and Allergy Thyroid Neoplasms Peroxidase Plant Proteins Chemiluminescence Detection limit biology medicine.diagnostic_test Chemistry Molecular biology Neoplasm Proteins Endocrinology Immunoassay biology.protein Alkaline phosphatase Soybeans Quantitative analysis (chemistry) |
| Zdroj: | Journal of Immunological Methods. 362:127-130 |
| ISSN: | 0022-1759 |
| Popis: | Serum thyroglobulin (Tg) is a main marker of thyroid cancer relapses after total or near-total thyroidectomy of patients with differentiated thyroid carcinoma. In this study, we developed a chemiluminescent enzyme-linked immunosorbent assay (CL-ELISA) for detecting Tg in human serum. Soybean peroxidase (SbP) in combination with 3-(10′-phenothiazinyl)propane-1-sulfonate (SPTZ) and 4-morpholinopyridine (MORPH) and horseradish peroxidase (HRP) with p-iodophenol (PIP) were used as detection systems in the sandwich CL-ELISA. Comparison of these two systems showed that a lower detection limit (LOD) of CL-ELISA with SbP/SPTZ/MORPH was 10 times lower than for the immunoassay with HRP/PIP. The LOD value for SbP-based CL-ELISA of 0.2 ng/mL was identical to LOD value typical of CL-ELISA Immulite® kit produced with alkaline phosphatase. The sensitivity of Tg CL-ELISA using SbP/SPTZ/MORPH completely satisfies the requirements of modern endocrinology. Comparative study of clinical serum specimens assayed by the SbP-based CL-ELISA (x) and Immulite® kit (y) for detecting Tg showed a good correlation between these two immunoassays (y = 1.15x − 0.14, R = 0.99). The obtained results open good perspectives for use of SbP/SPTZ/MORPH system in the development of ultra-sensitive immunoassays. |
| Databáze: | OpenAIRE |
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