Novel Ac Electrokinetic Platform For Rapid Isolation And Characterization Of Extracellular Vesicles From Nsclc Patients

Abstract We have developed a novel Alternating Current Electrokinetics (ACE) platform to isolate macromolecules such as circulating cell-free DNA (ccfDNA) and nanoparticles, including cell-derived extracellular vesicles (EVs). ccfDNA and EV isolation, and antibody staining using the platform required less than two hours to complete. EVs isolated by ultracentrifugation from the pancreatic cancer cell line ASPC-1 were used to validate the ACE platform's performance. EVs from pancreatic cancer ASPC-1 cells were detected using an anti-CD63 antibody and an antibody to the pancreatic cancer specific membrane protein Glypican-1, confirming that the ACE platform could isolate EVs and identify membrane-bound proteins. Recovered EVs were isolated and EV-associated RNA was amplified by RT-PCR. EVs from isolated from NSCLC patient plasma on the ACE platform contained higher levels of PD-L1 compared to healthy individuals (p<0.001). In parallel on the ACE platform, ccfDNA was isolated and quantified from the same samples, with increased levels measured in cancer patients. Improved differentiation between the NSCLC patients and the healthy normals was achieved when both markers were included in the analysis. Therefore, we believe the ACE platform has the potential to become a multimodal analytics tool for cancer diagnostics and treatment monitoring. Citation Format: Rajaram Krishnan, Juan P. Hinestrosa, David Searson, Robert Turner, Robert Kovelman, Heath Balcer, James Madsen, David Bodkin. Novel AC electrokinetic platform for rapid isolation and characterization of extracellular vesicles from NSCLC patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-174.