Development of ultra-sensitive soybean peroxidase-based CL-ELISA for the determination of human thyroglobulin

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• 作者：Marina M. Vdovenko ; Alex ; er V. Zubkov ; Galina I. Kuznetsova ; Leopoldo Della Ciana ; Nina S. Kuzmina ; Ivan Yu. Sakharov
• 关键词：Thyroglobulin ; ELISA ; Chemiluminescence ; Enhancement ; Peroxidase ; Soya
• 刊名：Journal of Immunological Methods
• 出版年：2010
• 出版时间：31 October 2010
• 年：2010
• 卷：362
• 期：1-2
• 页码：127-130
• 全文大小：312 K

文摘

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.