Novel Aptamer-Based Biosensor Platforms for Detection of Cardiomyopathy Conditions
Abstract
A point-of-care biosensing device for rapid screening and monitoring of cardiomyopathy conditions and progression to lower incidence death occurrences, and healthcare costs is needed. The aim of this project is to create a cardiomyopathy condition screening and monitoring tool to simplify the current biochemical marker testing procedures by developing vertically aligned platinum wire aptamer-based multi-array biosensor for precise, accurate, reliable, and rapid measurement of apt cardiac marker levels in the human whole blood and serum using electrochemical impedance spectroscopy. This work demonstrated construction of an impedimetric multi-array biosensor platform based on platinum wires functionalized with aptamers and successfully navigated the platform beginning from construction to optimization and validation of feasibility in biological samples. Initially, we optimized the optimal platinum wire diameter and surface finish necessary to create a biosensor that does not experience saturation within the acceptable clinical ranges of brain natriuretic peptide (BNP) and troponin T (TnT) antigens, the accepted cardiac biomarkers. Following validation, the focus shifted to assessing the self-assembled monolayer (SAM) approach utilized to tether the BNP and TnT specific aptamers to the electrode surface, determining both the optimal incubation time and concentrations needed for each layer and assessing the need of each layer. The best self-assembled-monolayer (SAM) combination that provided reliable, accurate and most sensitive response was determined as Platinum-Cysteamine-Glutaraldehyde-Neutravidin-Aptamer and this SAM combination showed excellent precision, reasonable sensitivity, and stable insulation of the linker proteins that easily interfere with the biosensor readings. The optimal SAM combination was also used to develop biosensors to test in human whole blood and serum samples to create a unique calibration curve model.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 01, 2023
- Accession Number
- AD1224425
Entities
People
- Abhijit Roy
- Mary Keebler
- Moni K. Datta
- Prashant N Kumta
- Sangeetha Kunjukunju
Organizations
- University of Pittsburgh