Developing a Biosensor-Based Immunoassay to Detect HPV E6 Oncoprotein in the Saliva Rinse Fluid of Oral Cancer Patients

• Published in: Journal of personalized medicine Vol. 12; no. 4; p. 594
• Main Authors: Cheng, Chi-Sheng, Ou, Bor-Rung, Lung, Feng-Di
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 07.04.2022; MDPI
• Subjects: Amino acid sequence; Antibodies; Antigens; Biomarkers; Biosensors; Carcinogenesis; Disease transmission; E6 protein; Enzyme-linked immunosorbent assay; Enzymes; Human papillomavirus; Immunoassay; Laboratories; Mass spectrometry; Medical research; Medical screening; Oncoproteins; Oral cancer; Oral cavity; Patients; Peptides; Polyclonal antibodies; Poultry; Precision medicine; Proteins; Saliva; Scientific imaging; Surface plasmon resonance; solid phase peptide synthesis; biosensor; surface plasmon resonance; human papillomavirus; E6 oncoprotein; salivary biomarkers; oral cancer
• ISSN: 2075-4426; 2075-4426
• DOI: 10.3390/jpm12040594

Transmission of Human papillomavirus (HPVs) is faithfully associated with carcinogenesis of oral cavity and oropharyngeal cancers. Therefore, clinical researchers may need to generate customized antibodies for the upcoming ELISA-based analysis to discover rare but valuable biomarkers. The aim of study was to develop and generate a biosensor-based immunoassay for early screening HPV-related oral cancer via saliva rinse fluid analysis. A peptide fragment of high-risk HPV subtype 16/18 protein, E6 protein (HP-1 protein sequence 48–66), was designed and synthesized, followed by the generation of polyclonal antibodies (anti-HP1 IgY) in our university-based laboratories. The titer and specificity of antibodies were determined by enzyme-linked immunosorbent assay (ELISA), and the Surface Plasmon Resonance (SPR) biosensor-based method was developed. Kinetic analyses by SPR confirmed that this designed peptide showed a high affinity with its generated polyclonal antibodies. Saliva fluid samples of thirty oral cancer patients and 13 healthy subjects were analyzed. SPR indicated that 26.8% of oral cancer patients had higher resonance unit (ΔRU) values than normal subjects. In conclusion, we developed a biosensor-based immunoassay to detect HPV E6 oncoprotein in the saliva rinse fluid for early screening and discrimination of HPV-related oral cancer patients.