Development of a Sol Particle Homogeneous Immunoassay for Measuring Full-Length Selenoprotein P in Human Serum

J Clin Lab Anal. 2016 Mar;30(2):114-22. doi: 10.1002/jcla.21824. Epub 2014 Dec 26.

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.

Keywords: colloidal gold; diabetes; homogeneous immunoassay; metformin; selenoprotein P.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anticoagulants / pharmacology
  • Calibration
  • Enzyme-Linked Immunosorbent Assay
  • Gold Colloid
  • Hemagglutination
  • Humans
  • Immunoassay / methods*
  • Kallikreins / blood
  • Limit of Detection
  • Proteolysis
  • Selenoprotein P / blood*

Substances

  • Anticoagulants
  • Gold Colloid
  • Selenoprotein P
  • Kallikreins