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

Xiaomei Yu,Shaoxiong Zheng,Yuan Qin,Liang Wang,Yigang Wang,Pengguo Xia,Biao Huang,Qiang He,Li Zhang,Xiumei Zhou,Xue Yang,Lingli Chen,Xiaobin Liu,Bo Lin,Zhigang Hu,Yueming Liu

doi:10.1007/s10895-022-02888-4

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 0.85ng/mL, and the detection range was 0.85-1000ng/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 ± 55.90ng/mL) compared to healthy controls (16.29 ± 9.91ng/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.

Highlights

Galactose agglutinin 3 (Galectin-3, galactose agglutinin 3 (Gal-3)), a member of β-galactoside-binding lectin family,[1] is an important regulators of inflammatory responses, and it is expressed in many inflammatory cells, such as macrophages.[2]
The Gal-3-time-resolved fluorescent immunoassay (TRFIA) was established using the double antibody sandwich method, with the capture antibodies coated on a 96-well microplate and the detection antibodies chelated to Europium (III) (Eu3+)
Serum Gal-3 was detected in 81 patients with Idiopathic Membranous Nephropathy (IMN) and 123 healthy controls to further evaluate the value of the Gal-3 in staging of IMN

Summary

Introduction

Galactose agglutinin 3 (Galectin-3, Gal-3), a member of β-galactoside-binding lectin family,[1] is an important regulators of inflammatory responses, and it is expressed in many inflammatory cells, such as macrophages.[2] When the cell is at rest, Gal-3 is stored in the cytoplasm; in the event of tissue damage, activated cells secrete Gal-3 via nonclassical pathways.[3] This protein binds to the cell surface and extracellular matrix polysaccharides and influences a variety of physiological and pathological processes, including apoptosis, adhesion, migration, fibrosis,[2] and inflammation.[4, 5] The expression of Gal-3 is detected in many disease states, such as renal disease.[6] Gal-3 is a biomarker associated with the progression of renal functional decline.[7] Elevated serum Gal-3 levels are associated with a rapid decline in glomerular filtration rate and an increased risk of chronic kidney disease (CKD).[8, 9] the role of Gal-3 in MN has not yet been reported

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Conclusion