Automated Multiplexed Electrochemiluminescence based Immunoarrays for Prostate Cancer Biomarker Protein Detection & Genotoxic Screening Assays Using Electrochemiluminescence and LC-MS/MS

Cancer is the leading cause of death in U.S. next to deaths caused by heart diseases. Cancer causes extreme pain to the patient physical and mental health. Prostate cancer is one of the most common type of cancer in American men besides skin cancer. Even though extreme efforts are being placed in the field of science and technology to develop reliable cancer therapeutics, no cure was found to eradicate or alleviate the pain caused by cancer. Cancer biomarker proteins that are abnormally expressed and secreted into blood in a diseased state could provide early cancer diagnosis and provide better healthcare. This helps in addressing better therapeutic options in much early stages of cancer even before advanced tumors developed where the survival rates are extremely low. As of now for prostate cancer, serum levels of prostate specific antigen is widely used as a diagnostic tool. But over expression of PSA could be from many other prostate related issues leading to false positives. This further results in causing patients stress due to exaggerated diagnosis and treatment options. Detection of panel of prostate cancer biomarkers from serum sample simultaneous with high sensitivity and selectivity could provide valuable information for early cancer diagnostics and better treatment options. The primary goal of this thesis is to develop ECL based automated diagnostic platforms that can detect panel of prostate cancer biomarkers with high sensitivity and high throughput. The assay platforms being low cost, rapid and non-complex they can be easily translated into public health care much faster and provide conclusive results with no ambiguity for better treatment options. This thesis explores many new device fabrication, automation strategies and liquid handing systems for easier completion of sensitive immunoassays with ultralow detection limits. Nano structured detection platforms and RuBPY dye doped silica nanoparticles have been used as amplification strategies to detect the serum proteins at low femtogram levels. We successfully developed immunoassay platforms that can detect small protein panel (3 proteins) to large protein (8 proteins) panel. We demonstrated 3-D printing as rapid fabrication tool to make microfluidic immunoarrays that enabled low cost sensors with relatively low sample volume requirements. The developed ECL arrays holds great promise in accurate detection of early stages of cancer in physician’s clinic and point of care settings