Circulating microRNAs (miRNAs) are a group of small RNA molecules that have been shown to be involved in several human diseases, such as cancers, neurodegenerative disease, and inflammation. Their sequences are highly conserved across species. miRNAs are present in biological fluids, are differentially expressed and can be used as biomarkers reported to be translational for research and drug development as improved diagnostic assays. Multiple techniques used to measure miRNA expression in biological fluids are labor-intensive and subject to variability. The objective of this study was to develop an extraction-free and amplification-free detection method using Single Molecule Counting (SMC™) technology and the SMCxPRO™ instrument combined with dynamic chemical labelling (DCL) chemistry by DESTINA for the direct detection of microRNAs in biological samples. As a proof of concept, miR-122 (miR-122-5p), a hepatocyte-specific microRNA biomarker for drug-induced liver injury (DILI) has been selected. A DESTINA a basic probe for detecting miR-122 is conjugated to superparamagnetic beads. These beads are added to DILI patient serums treated with DESTINA Stabiltech and incubated with a biotinylated DESTINA SMART Nucleobase. Following miR-122 target molecule hybridization to the DESTINA probe, the reactive SMART nucleobase is then covalently attached to the backbone of the probe by a dynamic covalent chemical reaction. The biotin-labeled superparamagnetic beads are then incubated with Alexa Fluor®647-conjugated streptavidin and the labeled heteroduplex complex is cleaved from the superparamagnetic beads before being detected by the SMCxPRO™ instrument. This is the first example where SMC™ technology has been used for molecular applications, allowing rapid quantification of circulating microRNA with single-based specificity and a limit of quantification suitable for research.
Speakers
Juan Díaz-Mochón
DESTINA
Chief Scientific Officer
Juan J. Díaz-Mochón is a chemical biology scientist turned biotech entrepreneur and promoter of the field of liquid biopsy as a way to improve patient care. He has enjoyed a distinguished academic career in Spain, Italy, and the UK, where he spent more than 10 years. Díaz-Mochón is the author of more than 70 peer-reviewed scientific publications and is the co-inventor of 14 patents. Building on one of his inventions, in 2010, he co-founded DESTINA Genomics Ltd. in Scotland with the ambition to become a leading manufacturer and supplier of custom chemical reagents and technologies for the best nucleic acid-based biomarker assays of his class. Díaz-Mochón, together with the late CEO of DESTINA, Hugh Ilyine, promoted the incorporation of the DESTINA subsidiary in Spain. Since then, DESTINA's R&D operations have been carried out in Spain, where cheaper, more accurate and much more significant nucleic acid testing tools are being developed by more than 10 full-time employees. Díaz-Mochón served as Chief Scientific Officer of DESTINA since its foundation and currently serves as Executive Director and CEO of DESTINA. In 2020, DESTINA, together with Roslin Technologies Limited, incorporated VETSINA Animal Diagnostics in Scotland, a company focused on developing and commercializing new veterinary assays. Díaz-Mochón is a member of the Scientific Advisory Committee of VETSINA. In the area of nonprofit organizations, in 2017, Díaz-Mochón co-founded the International Society for Liquid Biopsy, an organization committed to advancing Liquid Biopsy and promoting a multidisciplinary approach to the diagnosis and treatment of patients based on the use of liquid. biopsies. He is currently a Board Member of the Society. Díaz-Mochón holds a Ph.D. in Pharmacy from the University of Granada, studied as an undergraduate at the Università degli studi di Perugia and worked as a PDRA at the University of Southampton and the University of Edinburgh.
Webinar Information
SMC™ Technology
Duration:55 min
Language:English
Presented:
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