Self-assembled monolayers as the coating in a quartz piezoelectric crystal immunosensor to detect salmonella in aqueous solution

摘要

A new procedure based on the self-assembled monolayers (SAM) of alkanethiols was developed to immobilize antibodies onto gold electrodes of a quartz crystal microbalance (QCM) for detecting Salmonella paratyphi A. The procedure includes (1) chemisorption of 3-mercaptopropionic acid (MPA) at electrode surfaces to produce a carboxylic acid terminated monolayer, (2) activation by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydrosuccinimide (NHS) to generate a stable acyl amino ester intermediate, and (3) condensation of antibodies after aminolysis of the NHS adduct. Activation by two coupling agents (EDC and NHS) was shown to enhance the stability of the coating and facilitate the formation of a suitable intermediate to condense antibodies reproducibly and densely at the SAM, leading to high sensitivity and good precision of the developed immunosensor. With 50 min incubation under continuous monitoring, working ranges from 10(2) to 10(5) cells/mL with repeatability < 10% RSD were obtained. On the basis of S/N = 3, the detection limit was 1.7 x 10(2) cells/mL. The MO2 piezoimmunosensor developed was specific to differentiate S. paratyphi A against Escherichia coli and other serogroups of Salmonella that are commonly present together.