Quantitation without calibration : response profile as an indicator of target amount
Committee members: Baker, Gary M.; Hagen, Timothy J.; Horn, James R.||Advisor: Nesterova, Irina.||Includes bibliographical references.
Abstract
Quantitative assessment of biomarkers is essential in numerous contexts from decision-making in clinical situations to food quality monitoring to the interpretation of life-science research findings. However, appropriate quantitation techniques are not as widely addressed as detection methods. One of the major challenges in biomarker's quantitation is the need to have a calibration for correlating a measured signal to a target amount. The step complicates the methodologies and makes them less sustainable. In this work, we addressed the issue via a new strategy: relying on the position of response profile rather than absolute signal for assessment of a target nucleic acid's amount. To enable the capability, we developed a target-probe binding mechanism based on a negative cooperativity effect. A proof-of-concept example demonstrated that the model is suitable for quantitative analysis of nucleic acids over a wide concentration range. The general principles of the platform will be applicable for a variety of biomarkers such as nucleic acids, proteins, peptides, and others.
