B-212 Application of Annexin A5Coated Beads for Collection of Extracellular Vesicles and Their Content RNA as Liquid Biopsy for Lung Cancer

Abstract

Abstract Background Extracellular vesicles (EVs) carry molecules derived from cells, such as RNA. Measurement of tumor-derived extracellular RNA (exRNA) in EVs from plasma can be a promising approach for the development of blood-based cancer detection. However, the lack of an easy method for EVs separation limits its application. The aim of this study is to establish a procedure for EV collection and exRNA detection for non-small-cell lung cancer (NSCLC) diagnosis and monitoring. Methods Annexin A5-coated magnetic beads (ANX beads) were developed for EVs isolation. Next, we established a procedure for EV separation and exRNA detection from fluidic specimens using the ANX beads. Then blood samples from 22 NSCLC patients and 20 healthy controls were collected to evaluate the procedure and we detected different RNAs in the samples. Results We evaluated the ability of ANX beads in EVs separation and found that the beads could capture large and small EVs and could increase RNA concentration. Then we detected different RNAs in the samples and found that CK19 mRNA, MALAT1 lncRNA, and miR155 microRNA had good performance to differentiate NSCLC from healthy controls. Conclusion In conclusion, this assay protocol could separate EVs to enrich RNA concentration. The EVs separation employed was captured by magnetic beads which could be adopted into an automatic nucleic acid extract system to conveniently concentrate and extract nucleic acid from EVs.