Abstract 1930: Advancing cervical cancer early detection: A novel lateral flow immunoassay targeting p16INK4a
Authors: Samrin F. Habbani, Sayeh J. Dowlatshahi, Scott C. Bolton, Lucy T. Tecle, Lisa Flowers, Jacqueline C. Linnes, Sulma I. Mohammed
Journal: Cancer Research (2025)

Cervical cancer ranks as the fourth most prevalent cancer among women globally, with a stark disparity in disease burden and mortality between high-income and low- to middle-income countries (LMICs). Sub-Saharan Africa faces the highest incidence and mortality rates, primarily due to limited access to preventive measures such as vaccines, screening, early detection, and treatment. With approximately 90% of cervical cancer cases occurring in LMICs, there is an urgent need for effective, affordable screening solutions. This study addresses this critical gap by developing a cost-effective, easy-to-use point-of-care (POC) screening test. Specifically, we aim to create a lateral flow immunoassay (LFIA) to detect the overexpression of cyclin-dependent kinase inhibitor 2A (CDKN2A or p16INK4a), a key tumor suppressor protein overexpressed in cervical cancer. The LFIA is designed to be capable of detecting p16INK4a in precancerous and cancerous cells from cervical swab samples, thereby providing an accessible and efficient tool for cervical cancer screening in resource-limited settings. The developed POC LFIA utilizes antigen-antibody interactions. It has four key components: a sample pad, a conjugate pad, a reaction membrane, and an absorbent pad. Cervical swab samples applied to the sample pad rehydrate the dried conjugates, allowing them to interact with the analyte and migrate to the reaction zone, where a gold nanoparticle (AuNP)-conjugated antibody facilitates detection. Initial validation of p16INK4a levels in clinical swab samples was performed using an enzyme-linked immunosorbent assay, which revealed significant overexpression of p16INK4a in high-grade squamous intraepithelial lesions (HSIL), confirming its role as a biomarker for cervical precancerous conditions. Biolayer interferometry was used to select antibodies with optimal nanomolar to picomolar binding affinities. The chosen antibody was conjugated with AuNPs through passive adsorption, achieving a stable 100:1 molar antibody-to-AuNP ratio over seven days. The test and control lines were optimized using in-house dipsticks, with 1.5 mg/mL anti-mouse immunoglobulin G proving ideal for the control line. The current test can reliably detect p16INK4a concentrations as low as 0.5 μg/mL, with further refinements underway to lower the detection limit for clinical use. This simple and cost-effective LFIA offers robust performance, addressing the critical need for affordable and accessible cervical cancer screening, particularly in LMICs. With continued optimization, this tool holds significant promise for improving early detection and treatment outcomes in resource-limited settings. Keywords: point-of-care test, cervical cancer, p16INK4a, early detection, prevention, LMICs Citation Format: Samrin F. Habbani,Sayeh J. Dowlatshahi,Scott C. Bolton,Lucy T. Tecle,Lisa Flowers,Jacqueline C. Linnes,Sulma I. Mohammed. Advancing cervical cancer early detection: A novel lateral flow immunoassay targeting p16INK4a [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1930.