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The p16INK4a Antibody Immobilization Method for Immonosensor Application

  • Yang, Li (Frequency Control Laboratory, School of Automation & Engineering, University of Electronic Science and Technology of China) ;
  • Huang, Xian-He (Frequency Control Laboratory, School of Automation & Engineering, University of Electronic Science and Technology of China) ;
  • Sun, Liang (Department of Pathology, West China Second University Hospital, Sichuan University)
  • Published : 2015.07.13

Abstract

Background: The $p16^{INK4a}$ is a protein that expressed in Liquid-based cervical cytology specimens and has been proved link to cervical cancer. The $p16^{INK4a}$ could be detection by piezoelectric immunosensor and the immobilization of the $p16^{INK4a}$ antibody influence the sensitivity of the piezoelectric immunosensor. Materials and Methods: $5{\mu}L$ mouse polyclonal antibody against $p16^{INK4a}$ was bound onto the surface of immonosensor through two methods. (directly immobilized method; protein A method). Absorb of the $p16^{INK4a}$ antibody on the surface of immonosensor caused a shift in the resonant frequency of the immunosensor and The frequency changes recorded showed a better reproducibility. The activity of the immobilization antibody with the directly method and protein A method was tested with $p16^{INK4a}$ antigen. Results: The resonant frequency for different antibody immobilization methods were different, and the sensitivity for $p16^{INK4a}$ detection also different. Conclusions: The protein A method was found to be much more better than the directly method for the immobilization of the p16INK4A antibody on the gold electrode of the quartz crystal for cervical lesion detection. The Protein A method created more reproducible and stable immobilization antibody layers with p16INK4A antigen.

Keywords

$p16^{INK4a}$;cervical cancer;protein A;antibody immobilization

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