Research Project
8913590
? DESCRIPTION (provided by applicant): There is a critical need for a stabilization platform that can preserve biospecimen integrity, completely free of cold storage requirements. Such a technology would allow researchers, clinicians, and government personnel to conduct biomonitoring in field conditions (i.e. without requiring visits to the outpatient lab). Our long-trm goal is to develop a platform technology that maximizes the stability of a wide range of biospecimens in a novel silk protein matrix and that seamlessly integrates with downstream analytics. The objective is to evaluate the ability of a silk matrix to stabilize a number of bioanalytes, demonstrate integration of the silk system with several standard and emerging analytical techniques, and to prove the enhanced performance of silk-based protection to commercially-available solid stabilizers in accelerated aging studies. Our central hypothesis is that silk will comply with minimally- or non-invasive biospecimen collection routes, and provide comparable analyte retention to continuous frozen storage, while improving recovery as compared to commercially- available dried blood spot techniques. Our hypothesis has been formulated based on our preliminary data using silk to stabilize antibodies, plasma proteins, and nucleic acids from complex fluids such as whole blood, plasma, serum, and saliva. Therefore, in the first aim we will develop analytical methods to quantitate hazardous compound levels in plasma-laden silk matrices. This will be accomplished by incorporating techniques previously validated for use in analyzing dried blood spots. Modifications of silk molecular weight by proteolytic degradation can provide a route to remedy any potential negative impacts of the silk protein on downstream chromatographic techniques. As a second aim, we seek to enhance the recovery of hazardous compounds from silk matrices. For this aim, we will incorporate soluble additives to the silk protein reconstitution solutions to disable charge- or hydrophobic-mediated interactions that may retain the analyte prior to chromatographic separation. The expected outcomes would be a broader fundamental basis behind silk stabilization and new methods to translate these findings towards biospecimen recovery and storage as they pertain to biomonitoring. The rationale for the proposed research is that, once recovery and stability of these biomarkers is proven to be augmented by a silk encapsulate, diagnostic kits can be outfitted with various silk formats, which can then be used as a platform system for full validatio in follow-on studies in Phase II. Success with the planned approach can positively impact many areas of need in the field of diagnostics and epidemiology. These needs include alternative approaches at solid biospecimen stabilization, and translation/validation of analytical techniques important for hazard exposures. Furthermore, by offering methods to interface with finger-prick volumes (i.e. less invasive quantities) of whole blood, a silk-based stabilization system could significantly reduce financial burdens associated with the outpatient clinic, and in turn increase accessibility.
