BME PhD Preliminary Exam Announcement for Sayeh Jalali Dowlatshahi (J. Linnes, Advisor) Title: Quantification and Validation of Cervical Cancer-Associated Protein Biomarkers for Point-of-Care Early Cervical Cancer Detection Date: Tuesday, December 3, 2024 Time: 10:00 am Location: MJIS 2001 / Zoom Link: https://purdue-edu.zoom.us/j/8087054178<https://nam04.safelinks.protection.outlook.com/?url=https%3A%2F%2Fpurdue-edu.zoom.us%2Fj%2F8087054178&data=05%7C02%7Cbmeroundtable-list%40ecn.purdue.edu%7C5c7bbd94824a4baf9c5008dd07f537ab%7C4130bd397c53419cb1e58758d6d63f21%7C0%7C0%7C638675471956368726%7CUnknown%7CTWFpbGZsb3d8eyJFbXB0eU1hcGkiOnRydWUsIlYiOiIwLjAuMDAwMCIsIlAiOiJXaW4zMiIsIkFOIjoiTWFpbCIsIldUIjoyfQ%3D%3D%7C0%7C%7C%7C&sdata=33QCEoyLaf31ZYJg0WMKhNAQFb%2FjULTEL4EAHdm8S%2F8%3D&reserved=0> Thesis Committee Members: Prof. Jacqueline C. Linnes (Chair), Prof. Sulma Mohammed, Prof. Natalia Rodriguez, and Prof. Young Kim Abstract: Cervical cancer is the second most commonly diagnosed cancer in women and the second leading cause of cancer-related deaths among them globally. The incidence and mortality rates of this disease are considerably higher in low- and middle-income countries (LMICs). Current gold-standard cervical cancer screening techniques are expensive and time-consuming, possess low sensitivity or specificity, and require trained personnel and sophisticated instrumentation, limiting their implementation in low-resource settings. Therefore, a low-cost, rapid, sensitive, and specific point-of-care (POC) early-stage cervical cancer diagnostic assay is urgently needed. This research develops a complete workflow for cervical sample (1) collection and processing, (2) testing, and (3) result interpretation, for a POC cervical cancer screening test based on a paper-based lateral flow immunoassay (LFIA). Common cervical collection tools were compared to identify the one with optimal cellular recovery. Subsequently, a simplified cell lysis technique using radioimmunoprecipitation assay lysis buffer was devised for specimen processing. Four known key cervical precancer and cancer protein biomarkers were quantified in lysates of cultured cervical cells and clinical specimens using commercial enzyme-linked immunosorbent assays to assess their detection feasibility on paperfluidic platforms and obtain an estimate of their ranges, as these markers had only been studied qualitatively. The LFIA developed in this research detects topoisomerase II alpha (TOP2A), an enzyme innately involved in DNA replication that is overexpressed in cervical cancer patients. The test's colorimetric labels are 40-nm gold nanoparticles conjugated to mouse anti-TOP2A detector antibody molecules. Conjugation efficiency was evaluated through commercial quality control dipstick tests and dynamic light scattering. The chosen conjugate was then used with in-house dipstick assays to determine the control line's optimal goat anti-mouse immunoglobulin G antibody concentration. So far, the test line has detected as low as 50 ng/mL of TOP2A, showing promise that the platform will work in the 20-100-ng/mL clinical range. Upon optimization, these parameters will be incorporated into the TOP2A LFIA design. The device performance will be assessed with lysates of clinical cervical swab specimens. Furthermore, to streamline result interpretation, a machine learning (ML) classification algorithm will be developed based on the clinical samples' test line signal intensity and cytology category. This low-cost rapid test coupled with the cytology data-informed ML model will enable sensitive and specific cervical cancer screening in LMICs, leading to larger screening populations, more timely diagnosis, and earlier effective treatment, thereby significantly reducing the burden of this disease in those regions and worldwide. -- Bmeroundtable-list mailing list Bmeroundtable-list@ecn.purdue.edu https://engineering.purdue.edu/ECN/mailman/listinfo/bmeroundtable-list