Fluorescent Tools for Bioimaging & Industrial Process Development of Antibody-Drug Conjugates

 Fluorescence-based tools have revolutionized fundamental and applied research. Fluorescent tools  and techniques facilitate the drug discovery and development processes at various stages. Chapter  2 and 3 of this work are centered around the industrial process development of antibody-drug  conjugates, which are well-established anticancer therapeutics. The specificity of the monoclonal  antibodies towards cancer-cell surface antigen combined with the high potency of small-molecule  drugs in an ADC presents them as highly promising targeted therapeutics for cancer. They are  envisioned to grow rapidly in the next decade with numerous advancements in development of  superior conjugation technologies. However, the technical challenges and complexities associated  with manufacturing of antibody-drug conjugates impede these developments. This work aims to  address the barriers associated with hazardous ADC components and presents a solution for  expanding the ADC research in a safer and less-expensive way. Specifically, fluorescent non-toxic  surrogates of the most prominent warheads are designed to mimic the physicochemical properties  of the ADCs and facilitate their process development. Chapter 4 is focused on the fluorescent  peptide nucleic acids as FISH probes to label telomere-repeat containing RNA (TERRA). TERRA  is known to play a key role in alternative lengthening of telomeres in ALT-positive cancer cells  with low expression of telomerase enzyme. However, the exact mechanisms and associated  functions are unknown. The FISH PNA probes are shown to successfully label TERRA and have  potential application in live-cell imaging.