Luciferase-Based Detection of Antibodies for the Diagnosis of HPV-Associated Head and Neck Squamous Cell Carcinoma

Burbelo, Peter D.; Chaturvedi, Adrija; Notkins, Abner L.; Gunti, Sreenivasulu

Using the LIPS technology, antibody-based biomarkers against the human p53 tumor antigen Based on the substantial difference in antibody levels seen by LIPS between the HPV-negative and HPV-positive HNSCC samples, we sought to determine whether the rapid LIPSTICKS format, employing the NanoLuc-E6 fusion protein, could be used for detection of these antibodies in the HNSCC group ( Figure 2D ). The one-minute LIPSTICKS assays generated antibody signals that differentiated the HV and HPV-negative HNSCC patients from the HPV-positive HNSCC samples, in which the mean level of anti-E6 antibodies in the HNSCC samples was significantly higher (p = 0.03) than the value seen in the twenty HV controls ( Figure 2D ). Using a cutoff value based on the mean plus three standard deviations of the HV controls revealed a diagnostic performance of 89% (8/9) sensitivity and 100% specificity for detecting the E6 antibody-positive HNSCC patients in the rapid format. Moreover, the E6 antibody levels detected by the LIPSTICKS format were significantly different (p < 0.0001) between the HPV-negative HNSCC and HPV-positive HNSCC cases (data not shown). These findings suggest that the E6 LIPSTICK test has the potential to diagnose HPV-16-associated HNSCC.