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

Abstract

Point-of-care tests are needed for the screening of head and neck squamous cell carcinoma (HNSCC) and other malignancies. Luciferase immunoprecipitation systems (LIPS), employing light-emitting proteins, were used to examine serum antibodies against several cancer-associated targets in blood donor controls and subjects with colon cancer (CC) and HNSCC. The assessment of antibodies against the wild type p53 tumor antigen showed that approximately 25% of the CC and 20% of the HNSCC patients were seropositive. In addition, humoral responses against two p53 mutants, p53-R175H and p53-R273H, generally tracked the antibody responses seen against wild type p53. Analysis of antibodies against highly specific biomarkers of HPV-16-associated malignancy, E2, E6, and E7 oncoproteins, revealed no seropositivity in blood donors and CC patients. However, 45% (9/20) of the HNSCC patients showed E6 seropositivity, which overlapped all the detectable E2 (40%; 8/20) and E7 seropositive subjects (35%; 7/20). Using neodymium magnets, ultrarapid LIPSTICKS testing of HPV-16 E6 antibodies in <60 s per HNSCC sample demonstrated almost the same diagnostic performance (40% sensitivity and 100% specificity) as LIPS testing in 2.5 h. While additional improvements and standardization are needed, these results highlight the possibility of using these approaches for the diagnosis of HPV-16-associated HNSCC.

Beyond the Spike: identification of viral targets of the antibody response to SARS-CoV-2 in COVID-19 patients

“…Next, their ability to correctly identify COVID-19 patients was determined. Therefore, a cut-off value of LIPS antibody signals, based on the mean plus three standard deviations of the healthy negative control group (22)(23)(24), was calculated for each of these 11 antigens, along with the sensitivity and specificity of each test (Table 3, and Figure 4). The cut-off value of all 11 tests showed a high specificity of the SARS-CoV-2 LIPS assays with no samples from the negative control group being above the cut-off.…”

“…As many LIPS tests showed low sensitivity, we then used an approach based on the combination of LIPS antibody LU signals for the 11 separate SARS-CoV-2 antigen tests to efficiently detect SARS-CoV-2 exposure in this population (22)(23)(24). This approach of combining the LU values from the 11 LIPS tests increased the sensitivity to 100% for the diagnosis of COVID-19 infections during acute infection (Figure 5a, blue dots for <14 days).…”

Immunodiagnosis and Immunotherapeutics Based on Human Papillomavirus for HPV-Induced Cancers

“…Currently, many detection methods for the diagnosis and prediction of HPV-associated tumors are tried in basic research. For instance, a luciferase-based detection method for determining serum antibodies is possible for the diagnosis of HPV-associated head and neck squamous cell carcinoma ( 83 ). A new LSAB capture ELISA method based on the recombinant HPV16/18 E7 oncoproteins was used to investigate anti-HPV E7 antibody prevalence in the sera of patients with cervical cancer.…”

Screening for Porcine Deltacoronavirus in Human Serum Samples Via a Luminescent Immunoprecipitation System

“…Unlike enzyme linked immunosorbent assays (ELISAs), the highly sensitive LIPS is easily implemented to survey humoral serological response pro les to different antigens in a nearly universal format. It produces dynamic quantitative antibody titer ranges up to 6 log (10) for some antigens [32] [33,34]. LIPS also eliminates time and effort needed to produce highly puri ed bait antigens, optimization of host species speci c secondary antibodies, as well as the labor-intensive assay optimization steps needed for standard ELISAs [35].…”