The p16INK4a Antibody Immobilization Method for Immonosensor Application


Background: The p16INK4a is a protein that expressed in Liquid-based cervical cytology specimens and hasbeen proved link to cervical cancer. The p16INK4a could be detection by piezoelectric immunosensor and theimmobilization of the p16INK4a antibody influence the sensitivity of the piezoelectric immunosensor. Materials and
Methods: 5μL mouse polyclonal antibody against p16INK4a was bound onto the surface of immonosensor throughtwo methods. (directly immobilized method; protein A method). Absorb of the p16INK4a antibody on the surfaceof immonosensor caused a shift in the resonant frequency of the immunosensor and The frequency changesrecorded showed a better reproducibility. The activity of the immobilization antibody with the directly methodand protein A method was tested with p16INK4a antigen.
Results: The resonant frequency for different antibodyimmobilization methods were different, and the sensitivity for p16INK4a detection also different.
Conclusions:The protein A method was found to be much more better than the directly method for the immobilization ofthe p16INK4A antibody on the gold electrode of the quartz crystal for cervical lesion detection. The Protein Amethod created more reproducible and stable immobilization antibody layers with p16INK4A antigen.