1. Development of a Sol Particle Homogeneous Immunoassay for Measuring Full‐Length Selenoprotein P in Human Serum
- Author
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Masatoshi Nakagen, Kazuhiko Takahashi, Yoshiro Saito, Toshinari Takamura, Naoto Matsuyama, Takeshi Urabe, Hirofumi Misu, Shuichi Kaneko, Yuki Kita, Yumie Takeshita, Mutsumi Tanaka, Seiji Kato, Takehiro Kanamori, and Toru Nagano
- Subjects
0301 basic medicine ,Microbiology (medical) ,medicine.drug_class ,Clinical Biochemistry ,chemistry.chemical_element ,Enzyme-Linked Immunosorbent Assay ,Gold Colloid ,Monoclonal antibody ,03 medical and health sciences ,Limit of Detection ,Selenoprotein P ,medicine ,Humans ,Immunology and Allergy ,Immunoassay ,chemistry.chemical_classification ,Chromatography ,medicine.diagnostic_test ,Hemagglutination ,Biochemistry (medical) ,Public Health, Environmental and Occupational Health ,Anticoagulants ,Original Articles ,Hematology ,Molecular biology ,Medical Laboratory Technology ,030104 developmental biology ,chemistry ,Homogeneous ,Colloidal gold ,Calibration ,Proteolysis ,Particle ,Kallikreins ,Selenoprotein ,Selenium - Abstract
Background Selenoprotein P (SeP), a selenium-rich extracellular glycoprotein, is the primary selenoprotein in the plasma. SeP plays an important role in the maintenance of selenium levels in the peripheral tissues. We developed a new sol particle homogeneous immunoassay (SPIA) for measuring full-length SeP (FL-SeP) levels in the human serum. Methods We used colloidal gold particles coated with two types of anti-SeP monoclonal antibodies, one recognizing the N-terminal side domain of SeP and the other recognizing the C-terminal side domain. Results The assay range was 0.2–9 mg/l, and the linearity was excellent. The within-day and between-day coefficients of variation ranged from 0.73% to 2.24% and 0.45% to 1.11%, respectively. Serum samples (n = 200) were examined using the newly developed assay system (employing a Model 7070 Hitachi automatic clinical analyzer) and the conventional enzyme-linked immunosorbent assay. These two methods were compared using the Passing–Bablok regression analysis; the resulting regression equation and correlation coefficient were y = 0.940x + 0.165 and r = 0.954, respectively. Conclusions Our new SPIA assay is a fully automated homogeneous immunoassay that can be used in conjunction with various commercial analyzers. The assay was sensitive, precise, and suitable for clinical measurement of the FL-SeP in the human serum.
- Published
- 2014