Abstract
A kind of signal enhanced electrochemical biosensor based on double antibody sandwich film modified Au electrode was prepared by using layer-by-layer self-assembly technique. The Au electrode was firstly assembled with L-cysteine and glutaraldehyde, then fixed with c-Myc (9E10) monoclonal antibody (C-Ab1), resulting in forming a C-Ab1 modified electrode, which was able to recognise the c-myc oncogene protein. After combining with a second antibody of anti-mouse IgG (H+L) antibody (C-Ab2), the double antibody sandwich film (C-Ab1/c-myc/C-Ab2) modified electrode was obtained, in which the response signal was greatly enhanced, and the sensing property was better than that of the C-Ab1 modified electrode. The mechanism for signal enhancement with the double antibody sandwich method was explored through electrochemical impedance analysis and cyclic voltammetry behaviors. The impedance value of C-Ab1/c-myc/C-Ab2 modified biosensor was linearly proportional to the logarithmic concentrations of c-myc protein over the range of 0.043-430 nM, and the linear equation was fitted as <italic>Y</italic> = 10046.10 + 863.33<italic>X</italic> with a correlated coefficient of 0.9904. The detection limit of c-myc protein was evaluated to be 25.76 pM. The biosensor was simply prepared which possessed nice selectivity, reproducibility, stability and regenerability, and could be applied for determination of c-myc in rat serum with a recovery from 97.4% to 103.7%, indicating its practical application on detection of c-myc in tumor samples with potential value in biomedical field.
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