Measurement of recombinant human α-galactosidase A activity by enzyme-linked immunosorbent assay. The mouse anti α-gal antibody determined the degree of degrading α-gal-Bovine serum albumin according to the efficiency of recombinant human α-galactosidase A under various pH conditions. White circle, background; black circle, no enzyme; white square, enzyme in pH 8.0; white triangle, enzyme in pH 7.0; black triangle, enzyme in pH 6.0; black square, enzyme in pH 5.0.|@|~(^,^)~|@|Measurement of recombinant human α-galactosidase A activity using ion exchange column chromatography. (A-C) The α-gal synthetic type I was treated with recombinant human α-galactosidase A for 0 hour, 2 hours, and 6 hours. The output productions of monosaccharide which was cut by enzyme, were analyzed by high-pressure anion-exchange column chromatography. Galactose (E) and lactose (D) were used as controls.|@|~(^,^)~|@|Removal of α-gal epitope from porcine aortic endothelial cells (PAECs) by recombinant human α-galactosidase A. PAECs were treated with 10 U/mL recombinant human α-galactosidase A (dotted line) or mock treated (solid line). Griffonia simplicifolia 1-B4-fluorescein isothiocyanate was used to detect the presence of α-gal on the surface of PAECs using flow cytometry.|@|~(^,^)~|@|Removal of α-gal epitopes on the bovine pericardium by recombinant Bacterides thetaiotaomicron α-galactosidase using lectin histochemistry. Fresh bovine pericardium (D). Bovine pericardium treated with decellularization (E). Bovine pericardium treated with decellularization and α-galactosidase (F). (A), (B), and (C) were negative control of (D), (E), and (F), respectively. The frozen tissue samples were stained with biotinylated lectin and avidin-peroxidase to detect α-gal epitope. 3,3'-Diaminobenzidine staining intensity of tissues was captured by light microscopy (×400).|@|~(^,^)~|@|Quantitative evaluation of α-gal on the surface of bovine pericardium. The isolectin binding to fresh bovine pericardium (fresh), bovine pericardium treated with decellularization (D), and bovine pericardium treated with decellularization (D) and recombinant Bacterides thetaiotaomicron α-galactosidase (E), (D+E).|@|~(^,^)~|@|Transmission electron microscope (TEM) assessment of bovine pericardium (×400). TEM results for the fresh bovine pericardium (A), bovine pericardium treated with decellularization (B), and bovine pericardium treated with decellularization and recombinant Bacterides thetaiotaomicron α-galactosidase (C).

Korean J Thorac Cardiovasc Surg 2012;45:380~389 https://doi.org/10.5090/kjtcs.2012.45.6.380