Immunodiagnosis for cervical cancer using antibody-gold nanoparticle conjugate

S. Tapaneeyakorn, C. Thepthai, N. Apiwat, T. Dharakul
National Science and Technology Development Agency, TH

Keywords: cervical cancer, immunodiagnosis, monoclonal antibody, p16INK4A

Summary:

Cervical cancer is a leading cause of cancer deaths among women. It is most often caused by persistent infections with high-risk human papillomavirus (HPV). Since current screening programs are either complex or costly to run, the number of cervical cancer cases increases every year. Immunodiagnosis is an alternative choice of quick and reliable screening methods. One of the most promising biomarkers for cervical cancer is p16INK4A (p16). The over-expression of p16 is linked to the oncogenic transformation caused by persistent HPV infection. Here, monoclonal antibodies specific to p16 were produced by using hybridoma technology for immunodiagnostic purposes. The p16 was expressed in E.coli and purified by affinity chromatography. The protein was used for mouse immunization. Hybridomas that produce antibodies against p16 were selected by ELISA. The antibodies were purified by Protein A affinity column. The specificity and sensitivity of the antibodies were determined by Western blotting and ELISA method. The selected anti-p16 monoclonal antibody with highest sensitivity was conjugated to gold nanoparticles by non-covalent electrostatic van der Waals’ forces. Immunochromatographic (IC) tests by using conjugated anti-p16 monoclonal antibody and purified p16 were successfully performed for the first time. This method might be further used for early detection of cervical cancer.