4.4 Article

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

Journal

JOURNAL OF FLUORESCENCE
Volume 32, Issue 2, Pages 629-636

Publisher

SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s10895-022-02888-4

Keywords

Diagnosis; Fluoroimmunoassay; Galectin-3; Idiopathic membranous glomerulonephritis

Funding

  1. Social Development Fund of Zhejiang Province [LGF20H200008]
  2. Key Research and Development Project of Zhejiang Province [2020C03066]
  3. Scientific Research Projects of Jiangsu Provincial Health Commission [LGY201801]
  4. precision medicine key project of Wuxi health commission [J202001]
  5. Opening Project of Zhejiang Provincial Preponderant and Characteristic Subject of Key University (Traditional Chinese Pharmacology)
  6. Zhejiang Chinese Medical University [ZYAOXYB2019004]
  7. Chinese National Natural Science Foundation [82172336]

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The aim of this study was to develop a wide detection range Gal-3-TRFIA assay and explore the elevated Gal-3 concentration in serum of patients with IMN.
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.

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