Preparation of Peptide Biomarkers and Their Antibodies for the Development of Biosensor-/ Biochip-based Methods for Detecting HPV-related Biomarkers in Saliva of Oral Cancer Patients

• Main Authors: CHEN, GUAN-TING, 陳冠婷
• Other Authors: LUNG, FENG-DI
• Format: Others
• Language: en_US
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/r6hc4n

碩士 === 東海大學 === 化學系 === 106 === Over the past three decades, cancer has become the leading causes of death in Taiwan, and oral cancer ranks fifth in Taiwan’s top ten cancers. Among the risk factors of oral cancer, human papillomavirus (HPV) is a new target for recent research, the prognosis and treatment methods for HPV related oral cancer are different from those for the oral cancer caused by traditional risk factors. Therefore, this study is going to prepare the peptide biomarkers and their antibodies for the development of biosensor-/ biochip-based methods for evaluating the risk of patients with oral cancer who are at the high risk of HPV infection, and immediately prescribe a course of treatment to reduce medical costs. In this project, firstly, we designed a peptide fragment of HPV type 16 E7 protein (named HP-3), and synthesized it by solid phase peptide synthesis, followed by purification and analysis using RP-HPLC and characterization by MALDI-TOF MS. Secondly, HP-3 was used as an antigen for preparing polyclonal antibodies in chicken. The titer and specificity of anti-HP-3 antibodies were determined by Western Blot, kinetic and affinity analysis were determined by surface plasma resonance (SPR)-biosensor. Finally, SPR-based biosensor methods were developed for detecting HPV-related proteins or antibodies in saliva of oral cancer patient. Results demonstrate that we developed a non-invasive and convenient method for detecting HPV-related biomarkers in saliva collected from patients with oral cancer, and the peptide biomarker and SPR-methods have considerable potential for screening oral cancer patients with HPV infection.