Many biologics such as vaccines are temperature sensitive and must be stored and transported at refrigerated/freezing temperatures. This need for temperature-controlled storage and distribution is especially challenging in low-resource settings. The goal of this project is to develop a new processing method, light-assisted drying (LAD), to dehydrate biologics in preparation for long-term dry-state storage at ambient temperatures to reduce or eliminate the need for cold storage. This study explores strategies for further expediting LAD processing times and for the simultaneous processing of multiple samples.
Cold-chain storage can be challenging and expensive for the transportation and storage of biologics, especially in low-resource settings. Recent research has demonstrated that anhydrous preservation in a trehalose amorphous solid matrix offers an alternative to freeze drying for the preservation of biologics. We have previously described a new processing technique, light assisted drying (LAD), to create trehalose preservation matrices of small volume (40 μL) samples. LAD uses illumination by near-infrared laser light to selectively heat water and speed dehydration. In this study we apply the LAD technique to large volume samples (250 μL) that are more comparable to therapeutic doses.
Cold storage can be challenging and expensive for the transportation and storage of biologics. We are developing a new processing technique, light-assisted drying (LAD), to prepare biologics for anhydrous storage in a trehalose amorphous solid matrix. Nucleic acid nanoparticles (NANPs) are an example of new biological products that require refrigeration. DNA and RNA have emerged as building blocks for versatile biological drugs, called therapeutic nucleic acids (TNAs). NANPs have been developed to simultaneously deliver multiple TNAs and to conditionally activate TNAs and control their immunorecognition. The structural and chemical instability of NANPs over long-term storage at ambient temperatures is a challenge that may hamper broad use of this promising technology. In this work we apply the LAD technique to NANPs. NANPs suspended in a droplet of trehalose solution are irradiated with a near-IR laser to accelerate drying. As water is removed, the trehalose forms a protective matrix. The laser allows for careful control of sample temperature during processing. This is important as NANPs are thermally sensitive. In this study, RNA cubes (a type of NANP) were LAD processed and then stored for 1 month. Damage to LAD-processed NANPs was assessed after storage using gel electrophoresis and compared to unprocessed controls stored at 4°C. The thermal histories of samples were monitored during processing to determine the importance of temperature excursions on NANP viability after processing. The trehalose matrix was characterized using polarized light imaging to determine if crystallization occurred during storage, damaging embedded NANPs. These preliminary studies indicate that LAD processing can stabilize NANPs for dry-state storage at room temperatures.
Protein-based therapeutics have been developed to treat a range of conditions and assays use immobilized capture proteins for the detection of diseases. A challenge in the development of protein-based products is maintaining the protein in the folded state during processing and storage. The most common method of stabilizing proteins for storage is lyophilization. However, the freeze-drying process remains expensive and many proteins that are lyophilized must be refrigerated or frozen to maintain functionality. Cold storage strategies can be challenging for the transportation of protein-based products and can be difficult or impossible in low resource settings. Recent research has demonstrated that anhydrous, or dry state, preservation in a trehalose amorphous solid matrix offers an alternative to freeze drying for the preservation of biologics. We have previously described a new processing technique, light assisted drying (LAD), to create trehalose preservation matrices. LAD uses illumination by near-infrared laser light to selectively heat water and speeds dehydration of small volume (40 μL) samples. Low end moisture contents (EMC’s) are necessary for storage at supra-zero temperatures and this low water content must be uniform to insure successful long-term storage of embedded biologics. Our previous work has demonstrated the ability of LAD to reach EMCs necessary for storage at elevated temperatures. In this work, Raman spectroscopy is used to assess the trehalose distribution and water content across LAD processed samples. Results indicate that the water content of LAD process samples is uniform. LAD is a promising technique for processing biologics in preparation for anhydrous storage.
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