苏氨酸磷酸裂合酶电化学纳米免疫传感器的-研究.pdfVIP

ABSTRACT Novel electrochemical nano-gold particles biosensors for salmonella and shigella based on electrochemical detection analysis which could achieve automation detection, free the effects of sample color and turbidity, and simple equipments were needed; the efficiency of enzymatic reaction; and immunoassay which pretreatments such as enrichment and purification were needed were prepared for food safety. Covalent bonding, electrostatic adsorption and self-assembled were used to immobilize biomolecules onto the glassy carbon electrode combined the advantages of chitosan and gold nanoparticles. And we choose a anti-SpvC monoclonal antibody strain(Salmonella antigen preparation) which has the stongest affinity with purified protein SpvC (phosphothreonine lyase) from five anti-SpvC monoclonal antibody strains by Biomolecular interaction analyzer means (Octet ® biolayer interferometry) , the equilibrium dissociation constant KD is 3.85E-10M, the binding constant and dissociation rate constant Kdis and Kon are respective 3.73.E +03 L / mol s-1 and 9.62E-07 s-. A novel SpvC immunosensor based on double-layer gold nanoparticles has been developed, which the first layer of gold nanoparticles fixed on glassy carbon electrode by the chitosan, and the monoclonal antibody of SpvC was adsorbed; the second layer of gold nanoparticals was fixed by droping electrode with thionine-chitosan/NG-HRP complex. The assembly procedures were monitored by cyclic voltammetric and alter