Instrument-free yes/no quantitative analysis of molecular biomarkers

Northern Illinois University, Dekalb IL

Investigators

Abstract

Contemplating on the recent pandemics, the general public came to realize and appreciate the vital role of do-it-yourself diagnostic devices for disease control and management. Such devices report whether a unique pathogen-associate molecule (also known as a molecular biomarker) is found in a human. To extend the benefit of accessible molecular diagnostics to a wider range of diseases and situations, it is critical to develop devices that not only report whether a specific molecular biomarker is present but also answer the question of how much of that biomarker is present (so called quantitative analysis). The goal of this project is to develop a platform that enables an equipment-free and easy-to-interpret quantitative analysis of molecular biomarkers in do-it-yourself and point-of-care environments. To ensure an easy interpretation, the platform will produce a yes/no answer that involves observing bubbles as a readout. Observing bubbles does not require scientific training, an equipped lab, or color vision proficiency and, therefore, can be easily recognized by everyone ages 2 and up. In addition to public health benefits, the proposed development will spark and sustain a STEM interest in middle- and high school student through their direct hands-on engagement in the project- related experimental work. The goal of this project is to develop a platform for instrument-free easy-to-interpret quantitative analysis of molecular biomarkers. The proposed platform will comprise two developments: a yes/no output for quantitative measurement and a novel equipment-free signal readout. The yes/no quantitative measurement will be enabled through stoichiometry. The heart of the model is negative cooperativity-based target – probe binding. The binding modality yields a well-defined structure exactly at the stoichiometric equivalence point. Detection of the structure is a yes/no event for a quantitative result. The new equipment-free readout will be based on bubbling produced in a gas-generating reaction. As an easy to spot and interpret phenomena, bubbling perfectly matches the yes/no paradigm. The gas-generating readout will be triggered via an activatable in the equivalence point catalytic system. The project will produce a general methodology that is adaptable to a range of molecular targets including potential new agents (once their target binding is characterized to some extent). 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.