Establishment of Galectin-3 Time-resolved Fluoroimmunoassay and its Application in Idiopathic Membranous Nephropathy

J Fluoresc. 2022 Mar;32(2):629-636. doi: 10.1007/s10895-022-02888-4. Epub 2022 Jan 13.

Abstract

The aim of this study was to establish a time-resolved fluorescent immunoassay (TRFIA) for the detection of serum Galectin-3 (Gal-3) and apply this method to evaluate the clinical significance of serum Gal-3 in predicting Idiopathic Membranous Nephropathy (IMN) progression. The Gal-3-TRFIA was established using the double antibody sandwich method, with the capture antibodies coated on a 96-well microplate and the detection antibodies chelated with Europium (III) (Eu3+). Serum Gal-3 was detected in 81 patients with IMN and 123 healthy controls to further evaluate the value of the Gal-3 in staging of IMN. The sensitivity of the Gal-3-TRFIA assay was 0.85 ng/mL, and the detection range was 0.85-1000 ng/mL. The Gal-3 intra-batch and inter-batch coefficients of variation were 3.45% and 5.12%, respectively. The correlation coefficient (R) between the Gal-3-TRFIA assay and commercially available enzyme-linked immunosorbent assay kits was 0.83. The serum Gal-3 concentration was higher in patients with IMN (65.57 ± 55.90 ng/mL) compared to healthy controls (16.29 ± 9.91 ng/mL, P < 0.0001). In this study, a wide detection range Gal-3-TRFIA assay was developed using lanthanide (Eu3+) chelates for the detection of Gal-3 concentrations in serum. Gal-3 concentration is elevated in patients with IMN.

Keywords: Diagnosis; Fluoroimmunoassay; Galectin-3; Idiopathic membranous glomerulonephritis.

MeSH terms

  • Antibodies / blood
  • Disease Progression
  • Enzyme-Linked Immunosorbent Assay
  • Fluoroimmunoassay / methods*
  • Galectin 3 / blood*
  • Galectin 3 / immunology
  • Glomerulonephritis, Membranous / blood*
  • Glomerulonephritis, Membranous / diagnosis*
  • Humans
  • Prospective Studies
  • Sensitivity and Specificity
  • Time Factors

Substances

  • Antibodies
  • Galectin 3