title: Electrochemical assay of concanavalin A-ovalbumin binding on magnetic beads.
abstract: To monitor protein-glycoprotein interactions on magnetic beads, the present study developed an electrochemical assay of the binding between concanavalin A (ConA) and ovalbumin (OVA). The system was a powerful model that could be used to evaluate cell junctions. ConA with an electroactive daunomycin was immobilized on 6 different sizes of magnetic beads (diameter: 1.0-8.9 μm) through a cross-linking agent. Six sizes of OVA-beads (diameter: 1.0-8.9 μm) were also prepared using a similar method. The binding was evaluated using an oxidation peak of ConA with daunomycin because ConA recognized OVA with α-mannose residues. When binding took place on the beads' surface, the peak current was decreased due to the electroactive moieties being covered with OVA. When ConA/daunomycin-OVA binding was evaluated, the change of the peak current obtained by the beads (diameter: 8.9 μm) modified with ConA and daunomycin was the greatest in the presence of OVA-modified beads (diameter: 2.5 μm). In contrast, particle agglomeration was observed for the smallest beads (diameter: 1.0 μm) with ConA/daunomycin and OVA. The results suggested that ConA-OVA binding depended on the size of beads. Thus, this method could be applied to measure protein-glycoprotein interactions on the cell surface.
authors: Maebashi Institute of Technology, Gunma 371-0816, Japan. kzsuga@maebashi-it.ac.jp
authors: Kazuharu
authors: K
authors: Sugawara
authors: Asako
authors: A
authors: Yugami
authors: Toshihiko
authors: T
authors: Kadoya
authors: Hideki
authors: H
authors: Kuramitz
authors: Kohei
authors: K
authors: Hosaka
mesh_terms: D003208:Concanavalin A / Q000737:chemistry / Q000378:metabolism*; D003630:Daunorubicin / Q000737:chemistry; D004563:Electrochemistry / Q000379:methods*; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D059346:Magnets / Q000737:chemistry*; D008358:Mannose / Q000737:chemistry; D008863:Microspheres; D010047:Ovalbumin / Q000737:chemistry / Q000378:metabolism*; D010316:Particle Size; D011485:Protein Binding
chemical_list: D055767:Immobilized Proteins; D003208:Concanavalin A; D010047:Ovalbumin; D008358:Mannose; D003630:Daunorubicin