Room temperature biospecimen storage in aqueous environment is a critical requirement in National Cancer Institute’s (NCI) best practices for maximizing the quality of biospecimens. Presently, conventional techniques such as cryo or refrigeration-based storage techniques are employed. However, these techniques require complex matrix systems and is highly expensive making it impractical for many laboratories. Storage of biospecimens in their near native environment at room temperature can have a transformative global impact. However, this remains a challenge to date due to the rapid degradation of biospecimen over time. Here, we demonstrate an aqueous Capture and Release Gels for optimized storage (Bio-CaRGOS) of biospecimens (nucleic acids and proteins). We demonstrate 95% recovery of hemoglobin at 25°C (1-month) and 96 % recovery (7-months) at 4 °C. Hemoglobin is a marker of oxidative injuries, anaemia, hypertension, renal toxicity and regularly used in clinics applications i.e., blood donations, transfusion. The sterile bioCARGOS are achieved using a very low concentration of TMOS and water suspension that is hydrolyzed in standard microwave typically for 30 s and biospecimen of interest can be added to the hydrolysis at room temperature resulting in encapsulation. One major drawback of this method was generation of methanol during hydrolysis which is deleterious which would cause significant denaturation of the biospecimens. To address this critical issue, we distilled the CARGOS formulations and we achieved complete removal of methanol as indicated by Raman spectroscopy. Dynamic light scattering (DLS) experiments along with zeta potential studies suggests that the distilled samples are highly stable colloidal solutions. In conclusion, a novel, facile and low concentration aqueous CaRGOS have been formulated using a bench top microwave that allows for the long-term stabilization of biospecimens. Spectroscopic studies indicate complete hydrolysis of silica precursor that can be utilized to stabilize any biospecimen of interest based on its isoelectric point. Another challenging issue is the interference of silica in the downstream analysis which has been addressed via the use of inverted syringe filtration of encapsulated biospecimen that separates the CaRGOS and encapsulated biospecimen prior to downstream analysis. This approach significantly reduces the interference of silica in the downstream processing thereby giving more accurate and reliable data.

Bio-CARGOS: Capture and release gels for optimized storage of biospecimens at room temperature and its downstream applications

ACS Spring 2022

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