Multiplexed Hormone Test for Personalizing Menopause Treatment

Project Abstract Burst Diagnostics (BurstDX), in partnership with FemmeLoop, is developing a multiplexed, quantitative point-of- care and at-home diagnostic to measure estradiol (E2), progesterone (PG), luteinizing hormone (LH) and Follicle Stimulating Hormone (FSH). The diagnostic will measure levels in blood and saliva to enable determination of perimenopause/ menopause status and personalization of therapies including but not limited to hormone re- placement therapy (HRT). Perimenopause/menopause affects 27M women in the USA, impacting 20% of the workforce at a cost of $1.8B. Menopause treatment is growing, with the market for diagnostics and treatment currently at $17B. More frequent hormone level checks could be incredibly beneficial for patients and clinicians. However, the current regime for testing is 6-12 months. Consistent, timed daily to weekly tests are required to establish effective treatment but current laboratory testing cannot meet this demand limited by the high costs of testing ($80-$150). At-home multiplexed hormone detection represents an analytical challenge due to the small molecular size and low hormone concentrations in saliva and blood. E2 levels are as low as 1 pg/mL, progester- one levels as low as 10 pg/mL, LH levels as low as 0.5 mIU/mL, and FSH as low as 0.1 mIU/mL. Lateral flow assays (LFAs) and enzyme linked immunosorbent assays (ELISAs) are commercially available but don’t meet the demand. LFAs are easy to use but lack sensitivity and quantitative abilities. Moreover, hormone measure- ments necessitate the use of competitive immunoassays capable of detecting analyte at pg/mL levels, a feat that is virtually unattainable with LFAs. As a result, the market lacks any quantitative LFAs specifically designed for monitoring perimenopause/HRT. As a result, there is a clear need for a new at-home multiplexed test that is easy to use, low cost, accurate, and quantitative. Burst Diagnostics (BurstDX) is developing the Capillary-flow Driven Immunoassay (CaDI) as an alternative point-of-care ELISA that provides fast, specific, quantitative, multiplexed results. Our goal is to develop a reader that costs <$50 (for 100 runs) and assays that will cost ~$10/test. CaDI uses capillary flow-based microfluidics to perform all steps of a traditional ELISA including delivery of sample, enzyme-labeled detection antibody, washing buffer, and a chemiluminescent substrate for sensitive detection. Fluid flow is controlled by capillary wicking, eliminating pumps. The technology was licensed from Colorado State University with multiple published papers on the underlying concepts. BurstDX has improved sensitivity (using chemiluminescence), reproducibility, and manufacturability. We have demonstrated excellent performance for protein detection using sandwich immunoassays and have strong preliminary data for small-molecule competi- tive immunoassays. This proposal will expand testing into multiplexed immunoassays for E2, PG, LH and FSH in saliva and blood during Phase I to address the clear clinical and market need identified above before expand- ing in Phase II to develop the low-cost reader and perform clinical studies and usability testing.