New nanoparticle aided glycovariant biomarker tools to detect extracellular vesicles as a liquid biopsy for early diagnosis of bladder cancer

Abstract

Bladder cancer (BlCa) remains a significant health concern worldwide, with a high incidence rate and substantial morbidity and mortality. Early detection of bladder cancer is critical for timely intervention and improved patient outcomes. Therefore, there is an urgent need for non-invasive and reliable biomarkers that can enhance the accuracy and efficiency of bladder cancer detection. The aim of this project is to develop and validate highly sensitive nanoparticle-based time-resolved fluorometry immunoassay (TRFIA) to detect bladder cancer patients compared to clinically challenging benign samples. This project focused on the development of a combination of biotinylated antibody as a capture and europium chelate-doped nanoparticles (NPs)-coated lectin as a tracer used in immunoassay for the detection of BlCa patients. Captures such as cell adhesion molecules (EpCAM & CAM1) and cancer-associated integrin alpha-3 (ITGA-3) markers in combination with Ulex Europeaus Agglutinin (UEA) lectin were tested to find out the best functional biomarkers and their corresponding potential assays. Then the functional biomarker combinations were characterized and validated using cell culture medium (CCM) derived from cancer cell lines as well as pooled serum of benign and BlCa patients. We have observed fold changes with various salts and buffer optimization. Specifically, when employing UEA lectin, we obtained p-values of .03, .04 and .05 for EpCAM, CAM1 & ITGA-3 respectively. The outcomes of this project highlight the significance of systemically screening of UEA lectin with different captures to improve the development of a glycovariant assay for the detection of bladder cancer patients.