Advanced development and validation of aliquot-level visual indicators of biospecimen exposure to thawed conditions

/Abstract Many biological analytes of interest to both clinical oncologists and cancer researchers are unstable when the unfixed biospecimens in which they reside are exposed to thawed conditions. For example, American Society of Clinical Oncology / College of American Pathologists guidelines state that the post-excision-up-to-fixation exposure time span for tissues collected for clinical HER2 testing in breast cancer must be less than 1 hour. For blood plasma/serum and many types of tissue specimens that are to be frozen, the proper cold storage temperature is well below the common laboratory freezer temperature of -20 °C. For this and many other reasons, every year improprieties and inconsistencies in pre-analytical sample handling and storage generate unacceptably large numbers of costly false leads in biomedical research. Unsurprisingly, experts in the field agree that this problem must be minimized immediately. Currently there are few tools and not so much as one widely accepted approach by which to implement evidence-based tracking of biospecimen exposure to thawed conditions. In practice, it is actually quite rare for biomedical researchers to employ any evidence-based QA/QC tools at all—which suggests that easy-to-use, individual aliquot-level thawed-state indicators could have a major impact on improving biospecimen quality tracking and therefore actual biospecimen quality. In 2022, the PI’s lab was awarded an R21 grant through the NCI’s IMAT program to pursue development of the kinetically unique, autocatalytic, color-changing permanganate/oxalate reaction in melting point-depressed aqueous solvents as simple (i.e., user friendly), inexpensive, visual trackers of biospecimen exposure to inappropriately warm conditions, including temperature thresholds as low as -18 °C, -37 °C and -67 °C. The chemistry for 14 targeted time-temperature indicators (TTIs) has been successfully developed. Under this proposed project, the PI’s lab seeks to maximize the color intensity of the TTIs (advanced development) and to develop and validate user-friendly, self-contained, customer-activatable physical devices that will ultimately translate the unique chemistry that underpins the TTIs into market-ready products that provide aliquot-level color-changing indicators of biospecimen exposure to thawed or inappropriately warm conditions that make it essentially impossible for anyone handling the specimens to ignore their integrity status. This will be accomplished under two Specific Aims: Specific Aim 1: Leverage our recently developed, AI-enabled chemical kinetics simulations to develop and validate permanganate/oxalate reaction-based TTIs with maxed-out, 5x color intensity that run for as little as 5 minutes to as long as 2.5 hrs at 25 °C—and that have melting points of 0 °C, -18 °C, -37 °C, or -67 °C. Specific Aim 2: Develop and performance-test several physical device designs that will provide self-contained, aliquot-level, user-activatable, color-changing TTI devices that anyone can use to track the exposure of biospecimens to thawed / inappropriately warm conditions. Project Number: 1R33CA309724-01 | Fiscal Year: 2026 | NIH Institute/Center: National Cancer Institute (NCI) | Principal Investigator: CHAD BORGES | Institution: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS, SCOTTSDALE, AZ | Award Amount: $353,601 | Activity Code: R33 | Study Section: Special Emphasis Panel[ZRG1 BTC-V (55)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11312119