Electrochemical Biosensor : Point-Of-Care for Early Detection of Bone Loss

This book presents the design of a robust, portable and low-cost PoC sensing system for the early detection of bone loss. The device can measure the level of CTx-I – one of the most sensitive biochemical markers of bone resorption – in serum and transmit the measured value to an IoT-based cloud server. The selectivity of the sensing system to CTx-I has been achieved by coating the sensor with artificial antibodies, prepared by means of molecular imprinting technology. Explaining all aspects of the system’s development in detail, the book will be of great interest to all engineers, researchers and scientists whose work involves the development of electrochemical sensors and PoC devices. Preface 6 Contents 8 1 Introduction 11 Abstract 11 1.1 What Is Osteoporosis? 11 1.2 Osteoporosis Facts and Statistics 11 1.3 Osteoporosis Diagnosis 13 1.4 The Aim of the Book 14 1.5 Research Contributions 15 References 16 2 State-of-the-Art of Sensing Technologies for Monitoring of Bone-Health 17 Abstract 17 2.1 Introduction 17 2.2 Bone Structure 18 2.3 Biochemical Markers of Bone Turnover 18 2.3.1 Biochemical Markers of Bone Formation 19 2.3.2 Biochemical Markers of Bone Resorption 20 2.4 Analytical Methods for the Measurement of Bone Turnover Markers 23 2.4.1 Enzyme-Linked Immunosorbent Assay 23 2.4.2 Radioimmunoassay 23 2.4.3 High-Performance Liquid Chromatography 25 2.5 Current Advancements in Bone Biosensors 25 2.5.1 Biomechanical Sensors 27 2.5.2 Biomarker-Based Sensors 31 2.5.3 Multiplex Assays 33 2.6 Conclusions 35 References 36 3 Planar Interdigital Sensors and Electrochemical Impedance Spectroscopy 42 Abstract 42 3.1 Operating Principle of Interdigital Sensors 42 3.2 Novel Planar Interdigital Sensors 44 3.3 Electrochemical Impedance Spectroscopy (EIS) 47 3.4 Experimental Setup 49 3.5 Conclusions 51 References 52 4 Antigen-Antibody-Based Sensor for CTx-I Detection 54 Abstract 54 4.1 Introduction 54 4.2 ELISA-Based Experiments 54 4.2.1 Materials and Chemicals 55 4.2.2 Assay Procedure 55 4.2.3 Results 57 4.3 Ag-Ab-Based Biosensor 57 4.3.1 CTx-I Measurement in Known Samples 59 4.3.2 Data Analysis Using Non-linear Least-Square Curve Fitting 59 4.3.3 Multivariate Chemometric Analysis 62 4.3.4 CTx-I Measurement in Unknown Samples Using the Ag-Ab-Based Biosensor 62 4.4 Conclusions 64 References 65 5 MIP-Based Sensor for CTx-I Detection 67 Abstract 67 5.1 Introduction 67 5.2 General Principle of Molecular Imprinting Technology 68 5.2.1 MIP Categories 69 5.2.2 Effects of Monomers, Cross-Linker, Porogenic Solvents and Initiator in MIP 69 5.2.3 Preparation Methods of MIP 72 5.3 Materials and Methods 73 5.3.1 Materials and Apparatus 73 5.3.2 Preparation of Artificial Antibodies Using Molecular Imprinting Technology 74 5.3.3 Preparation of the Functionalised Biosensing Surface 74 5.3.4 Preparation of the CTx-I Samples 75 5.3.5 Experimental Measurements 75 5.4 Results and Discussions 76 5.4.1 SEM Characterisation 76 5.4.2 Sorption Studies of CTx-I to MIP and NIP 76 5.4.3 EIS Measurement and Analytical Measurement 78 5.4.4 CNLS-Based Biosensor Response 81 5.4.5 Measurement of CTx-I in Real Serum Samples Using the CNLS–Based Calibration Curve 82 5.4.6 Single-Frequency Reactance-Based Biosensor Response 83 5.4.7 Measurement of CTx-I in Real Serum Samples Using Single-Frequency-Based Calibration Curve 84 5.4.8 Comparison Between the CNLS-Based Measurement and Single-Frequency Measurement 84 5.5 Performance Assessment of Interdigital Sensor for Varied Coating Thicknesses 84 5.5.1 Coating the Sensing Surface with Acrylic 86 5.5.2 Coating the Sensing Area with Selective Material 86 5.5.3 CTx-I Measurement Using the Coated Interdigital Sensor 87 5.5.4 Sensitivity of the Coated Sensor and the Saturation Level 89 5.5.5 Dependence of Coating Thickness on Withdrawal Speed and Dipping Time 90 5.6 Conclusions 94 References 95 6 IoT-Enabled Microcontroller-Based System 100 Abstract 100 6.1 Introduction 100 6.2 Electrochemical Impedance Monitoring Using the Microcontroller-Based System 101 6.2.1 Block Diagram of the Point-of-Care System 101 6.2.2 Circuit Diagram of the Proposed System 102 6.2.3 Software Process Flow of the PoC Device 103 6.3 Results and Discussion 105 6.3.1 Calibration Curve Development from the Proposed System 105 6.3.2 Unknown Sample Measurement 106 6.3.3 IoT Data from the PoC Device 108 6.4 Conclusions 108 References 109 7 Summary and Conclusions 111 Abstract 111 7.1 Conclusions 111 7.2 Future Possibilities 112 References 114 Front Matter ....Pages i-ix Introduction (Nasrin Afsarimanesh, Subhas Chandra Mukhopadhyay, Marlena Kruger)....Pages 1-6 State-of-the-Art of Sensing Technologies for Monitoring of Bone-Health (Nasrin Afsarimanesh, Subhas Chandra Mukhopadhyay, Marlena Kruger)....Pages 7-31 Planar Interdigital Sensors and Electrochemical Impedance Spectroscopy (Nasrin Afsarimanesh, Subhas Chandra Mukhopadhyay, Marlena Kruger)....Pages 33-44 Antigen-Antibody-Based Sensor for CTx-I Detection (Nasrin Afsarimanesh, Subhas Chandra Mukhopadhyay, Marlena Kruger)....Pages 45-57 MIP-Based Sensor for CTx-I Detection (Nasrin Afsarimanesh, Subhas Chandra Mukhopadhyay, Marlena Kruger)....Pages 59-91 IoT-Enabled Microcontroller-Based System (Nasrin Afsarimanesh, Subhas Chandra Mukhopadhyay, Marlena Kruger)....Pages 93-103 Summary and Conclusions (Nasrin Afsarimanesh, Subhas Chandra Mukhopadhyay, Marlena Kruger)....Pages 105-108