Leprechaun

Importance of the Topic

Growing demand for precise quantification and characterization of virus vectors and extracellular vesicles makes robust, sensitive analysis crucial in bioprocess development and quality control.

Objectives and Study Overview

This study introduces Leprechaun, an integrated system designed to perform comprehensive biophysical and biochemical profiling of lentivirus particles and exosomes in crude or purified preparations with minimal sample preparation.

Methodology and Instrumentation

Leprechaun employs a proprietary Luni consumable for sample capture and analysis in three main steps:

• Capture on Luni surface using specific antibodies
• Size measurement via interferometric reflectance microscopy
• Immunofluorescence detection of structural and phenotypic markers

Used Instrumentation

The Leprechaun system features a cooled scientific CMOS sensor, IRM illumination at 415 nm, fluorescence LEDs at multiple wavelengths, and automated analysis software. It accepts sample volumes from 1 to 25 µL and processes up to 16 consumables per run.

Main Results and Discussion

• Lentivirus analysis resolved particle concentration, capsid presence, RNA content, and aggregate levels across different production stages, identifying high quality batches early
• Contaminant profiling distinguished enveloped vesicle impurities and quantified soluble p24
• Exosome profiling provided total counts, size distribution from 35 to 200 nm, and phenotype based on colocalization of up to four markers such as CD9, CD63, and CD81

Benefits and Practical Applications of the Method

• Rapid, multiplexed characterization with low sample consumption
• Elimination of extensive purification steps
• Real time process monitoring for biomanufacturing and academic research

Future Trends and Potential Applications

Anticipated developments include expanded marker panels, integration with automated workflows, extended dynamic range, and advanced data analytics for real time decision support in cell and gene therapy and diagnostic exosome studies.

Conclusion

Leprechaun delivers an all in one solution for accurate titer, structural and phenotypic profiling of lentivirus and exosomes, enhancing quality control and accelerating process development in virology and extracellular vesicle research.