Although frozen storage is currently the standard practice for storing of biomolecules used as reagents, the process is both inefficient and costly. To ensure stability, reagents must be stored at high concentrations and volumes resulting in significant material discard. While lyophilization can minimize some of these issues; lyophilization requires extensive regent-specific development, may result in loss of yield, and cannot be applied for materials that cannot tolerate freezing. To address these, we have developed a novel and efficient bio-preservation method called capillary-mediated vitrification (CMV). The CMV process is simple to perform at the bench and does not require reagent specific optimization, freezing step and presents minimal risk of matrix interference. The preservation process can be completed in under an hour, producing a stable, dried product. The approach is broadly applicable to different types of biomolecules including enzymes, protein conjugates, antibodies, nucleic acids, small molecules, and viruses. In this report, we used the CMV process to prepare a variety of reagents in a pre-diluted, single use format including: monoclonal antibodies and conjugates, luciferase and luciferin, mRNA and PCR reagents. CMV-preserved alkaline phosphatase and horse radish peroxidase conjugated human IgG antibodies incubated at 55°C overnight showed a similar signal of -20°C stored liquid antibody in an ELISA. The CMV stabilized luciferase and luciferin both maintained &gt;70% activity after storage at 45°C for 6 weeks. A CMV stabilized green fluorescent protein-expressing mRNA stored at 55°C for 100 days had similar transfection efficiency to mRNA that was stored at -80°C prior to use. The CMV-process efficiently preserved PCR reagents which also maintained activity when stored at elevated temperature. In conclusion, CMV is a promising alternative to traditional biopreservation methods and could significantly improve analytical workflows and reduce the need of cold storage for biological molecules used as reagents.

Capillary mediated vitrification: A novel approach for preparing stabilized, ready-to-use reagents

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ACS Fall 2022

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