This Small Business Innovation Research (SBIR) Phase I project fills a key technological gap in development of biological and chemical sensor development. There is an unmet need for tests that easily detect molecular, cellular, or clinical responses that indicate disease onset or predict which candidate drugs will have harmful side effects. In this project, we are addressing this need by innovating and commercializing a reliable, sensitive, and accurate test that simultaneously monitors and quantifies levels of multiple proteins in biological fluids (plasma, urine, serum). These target protein biomarkers indicate disease onset, disease progression, or drug efficacy. The test is simple and can be performed using existing laboratory equipment, allowing for easy adoption in academic labs and pharmaceutical companies. Furthermore, this test is the only test that reliably prevents false positive signals, which typically result from nonspecific binding between non-target protein-detection agents. Preventing false positive signals is crucial because it enables more accurate measure of proteins in biological fluids. An initial test panel of proteins to be tested will be for cardiovascular disease management and drug development.
The broader impact/commercial application of this project is in the $1 billion life science research market that today is served mainly by tests that can only detect a single protein at a time. Measuring proteins one by one is time- and labor-intensive. The proposed research has tremendous potential as a research tool for highly accurate detection and analyses of protein biomarkers, leading to better understanding of disease pathogenesis and therapeutic response. The innovative, new test also has potential applications in the $36.6 billion contract research organization market to improve drug safety and efficacy testing, decrease clinical trial costs, and decrease time to market. This proposal advances the fields of Biological and Chemical Technologies (BC) as well as of Nanotechnology, Advanced Materials, and Manufacturing (NM)
This Phase I Small Business Innovation Research project entailed development and prototyping of a test platform for detection of proteins involved in cardiovascular disease. The funded company, PHASIQ, has technology that enables the creation of multiple protein tests (multiplex immunoassays) in which unwanted molecular interactions (antibody crosstalk) are eliminated. This allows for the creation of multiplex immunoassays that analyze multiple proteins that are otherwise unattainable due to antibody crosstalk. The overall goal of this project was to create a prototype crosstalk-free multiplex immunoassay kit in which most reagents are pre-spotted and dehydrated on test plates. A user would need only add sample (which would rehydrate the immunoassay reagents) to begin the analysis protocol. The first objective of this project was focused on the development and fabrication of protein test wells, or a microwell plate, this is compatible with PHASIQ’s technology. Different geometric designs and materials were tested using 3D printing technology, by-hand fabrication methods, and injection molding. Preliminary experiments that investigated test chamber design were focused on ensuring reagent placement on plates was adequate and also allowed for solution dehydration without unwanted mixing. In addition to being compatible with PHASIQ’s technology, the custom plate was designed to meet potential customer and market demands. Work on this objective yielded a 32-well test plate that was used in the development of a working prototype for cardiovascular disease-indicating protein detection. More precisely, the test plate designed in the first objective was used to develop a manufacturing formulation for a user-friendly, disease-focused multiplex immunoassay kit. This kit requires minimal preparation from the end user, and has all of the immunoassay components pre-applied to the test plate. A user only needs to add sample, incubate, wash, and perform minimal detection procedures. To accomplish development of a tool with this capability, it was necessary to test dehydration, rehydration, and use of antibodies for immunoassays on plate developed in the first objective. A design of experiments (Taguchi) was used to optimize use of reagents and procedure for producing pre-spotted plates. Work towards the second objective resulted in a functioning dehydrated multiplex immunoassay plate created with PHASIQ’s unique technology. The prototype developed by completing this project is a useful tool for cardiovascular disease researchers with potential for eventual clinical diagnostic use.
