Sbir phase II: enzyme-linked immunosorbent assay for detection of per- and polyfluoroalkyl substances in water

Aqua Science LLC received a $1.23M Cooperative Agreement award from the National Science Foundation's Technology, Innovation, and Partnerships program (CFDA 47.084) effective June 1, 2025, through May 31, 2027. This Phase II SBIR (Small Business Innovation Research) project focuses on developing and commercializing an enzyme-linked immunosorbent assay (ELISA) for the rapid detection of per- and polyfluoroalkyl substances (PFAS) in water. The ELISA technology will serve as a cost-effective alternative to current liquid chromatography-mass spectrometry (LC-MS) testing methods, reducing per-sample costs from $250–$400 to approximately $40 while increasing daily sample throughput from 30 to approximately 120 samples.

The project's primary deliverable is a market-ready PFAS ELISA detection assay that addresses a critical national need, as PFAS contamination affects more than 95 percent of the U.S. population and has been linked to serious health conditions including thyroid disease and certain cancers. Development activities include creation of novel antibodies, hapten design, immunogen preparation, and antibody screening to achieve a two-orders-of-magnitude improvement in detection sensitivity over Phase I results. The resulting technology is intended to enable rapid on-site PFAS screening and assessment of remediation treatment efficacy for contaminated sites, directly supporting public health and environmental protection initiatives aligned with NSF TIP's mission to advance breakthrough technologies addressing national challenges.

Abstract

The broader impact of this small business innovation research (sbir) phase II project is in bringing an elisa (enzyme-linked immunosorbent assay) for the detection of PFAS (per- and polyfluoroalkyl substances) to market. Rapid, on-site testing using elisa would be an attractive low-cost solution for PFAS screening to complement current slow and costly LC-MS (liquid chromatography?mass spectrometry) method. As a result of PFAS? Pervasiveness, more than 95 percent of the U.S. Population has PFAS in their bodies. These compounds have been associated with high cholesterol levels, thyroid disease, certain cancers, and pregnancy-related problems. Since the amount of testing for PFAS is expected to increase over time, there is a significant need for a cost effective, rapid method for detection of PFAS. Today, an LC/MS sample test costs $250-$400 per sample and the throughput is low at 30 samples per day whereas an elisa test could be about $40 per sample and can test about 120 samples per day. Thus, an elisa assay for PFAS detection would be a groundbreaking technology for the testing to identify PFAS hotspots, and assessment of remediation treatment efficacy for many PFAS projects and clean UPS. This project thus aligns with the goal of promoting well-being of U.S. Population. Development of a PFAS elisa requires the use of an antibody. Currently no such antibodies are available commercially. Elisas are preferred due to their sensitivity and ability to tolerate pretreatments. The process of elisa development includes development of critical components such as hapten development, immunogen preparation, antibody production, screening of antibodies for specificity to varying PFAS as well as sensitivity. The intellectual merit of this phase II project lies in improving the PFAS detection sensitivity by 2-orders of magnitude compared to what was demonstrated in phase i. While phase i proved that PFAS can be detected using an elisa assay with a hapten for immunogen preparation in a single animal species, phase II will attempt to find antibodies produced by multiple animal species, specifically those that are not physically aggregated. The r & d focus initially will be on hapten and antibody- horseradish peroxidase (HRP) conjugate production. This will be followed by identification of the most sensitive pair and external validation of the test kits. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the foundation's intellectual merit and broader impacts review criteria.- subawards are not planned for this award.