Molecularly Imprinted Polymers: Methods and Protocols (Methods in Molecular Biology, 2359)

✍ Scribed by Antonio Martín-Esteban (editor)

• Publisher: Humana
• Year: 2021
• Tongue: English
• Leaves: 291
• Category: Library

✦ Synopsis

This detailed volume explores molecularly imprinted polymers (MIPs), which have attracted great interest both in fundamental research and for practical applications due to their selective molecular recognition capabilities, extraordinary stability, and ease of preparation. Beginning with key laboratory protocols describing the different steps towards the synthesis of MIPs by different polymerization strategies, the volume continues by examining MIP use in sample preparation, their implementation on the development of sensors, as well as applications in other areas such catalysis and the use of bioinformatics and molecular modeling for MIPs design. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to the respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Molecularly Imprinted Polymers: Methods and Protocols serves as an ideal guide for researchers seeking to harness this very powerful approach for the preparation of molecular selective synthetic polymers.

✦ Table of Contents

Preface
Contents
Contributors
Chapter 1: Synthesis of Molecularly Imprinted Polymers by Two-Step Swelling and Polymerization
1 Introduction
2 Materials
2.1 Reagents
2.2 Preparation of Polystyrene Seed Particles
2.3 Preparation of MIPs for (S)-Nilvadipine
2.4 Preparation of RAM-MIPs for BPA-d16
3 Methods
3.1 Separation of Nilvadipine Enantiomers
3.2 Simultaneous Determination of Bisphenol A and Its Halogenated Derivatives in River Water
4 Notes
References
Chapter 2: Molecularly Imprinted Polymeric Nanoparticles by Precipitation Polymerization and Characterization by Quantitative ...
1 Introduction
2 Materials
2.1 Polymerization
2.2 Template Extraction
2.3 Quantitative 1H NMR
3 Methods
3.1 Polymerization
3.2 Template Extraction
3.3 Quantitative 1H NMR Analysis
3.3.1 Determination of Template Incorporation and Polymer Composition (See Note 14)
3.3.2 Binding Tests (See Note 20)
4 Notes
References
Chapter 3: MIP Synthesis and Processing Using Supercritical Fluids
1 Introduction
2 Materials
2.1 MIP Synthesis in scCO2
2.2 ScCO2-Assisted Template Desorption
2.3 ScCO2-Assisted MIP Impregnation
2.4 ScCO2-Assisted Membranes Preparation
2.5 Applications
2.5.1 Drug Delivery MIPs
In Vitro Drug Release Experiments
2.5.2 MIPs for Purification Processes
Static Binding Tests
Solid Phase Extraction (SPE) Experiments
2.5.3 ScCO2-Assisted Preparation of a PMMA Composite Membrane
3 Methods
3.1 MIP Synthesis in scCO2
3.2 ScCO2-assisted Template Desorption
3.3 ScCO2-Assisted MIP Impregnation
3.4 ScCO2-Assisted Membranes Preparation
3.5 Applications
3.5.1 Drug Delivery MIP (See Note 6)
In Vitro Drug Release Experiments
3.5.2 MIP for Purification Processes
Acrylate and Acrylamide-Based MIPs for a Model Pharmaceutical Impurity (See Note 8)
Static Binding Tests
Solid Phase Extraction (SPE) Experiments
MIP-Layered Silica Beads for a Model Pharmaceutical Impurity (See Note 10)
Gravity-Driven Column Experiments
3.5.3 ScCO2-Assisted Preparation of a PMMA Composite Membrane
4 Notes
References
Chapter 4: Synthesis of Bacteria Imprinted Polymers by Pickering Emulsion Polymerization
1 Introduction
2 Materials
2.1 Stock Solutions
2.2 Chemical Synthesis
3 Methods
3.1 Preparation of Microorganism Samples
3.2 Synthesis of N-acrylchitosan (NAC) Pre-polymer
3.3 Synthesis of Bacteria-Imprinted Polymer (BIP) Beads
4 Notes
References
Chapter 5: Restricted Access Molecularly Imprinted Polymers
1 Introduction
2 Materials
2.1 RAMIP Synthesis: Hydrophilic Monomer Grafting
2.2 Template Removal from RAMIP
2.3 Epoxide Ring Opening
2.4 Coating with Bovine Serum Albumin
2.4.1 Covering the MIP Using a Solid Phase Extraction Cartridge
2.4.2 Covering the MIP Using a Glass Tube
2.5 Protein Exclusion Test
2.5.1 In Column Packed with RAMIP
2.5.2 In Solid Phase Extraction Cartridges
2.5.3 For Solid Phase Microextraction RAMIP Fibers
3 Methods
3.1 RAMIP Synthesis: Hydrophilic Monomer Grafting
3.1.1 RAMIP Obtained with Hydrophilic Monomer in a Single-Step Synthesis
3.1.2 RAMIP Obtained with Hydrophilic Monomer Added After a Pre-polymerization Step
3.2 Template Removal from RAMIP
3.3 Epoxide Ring Opening
3.4 Coating with Bovine Serum Albumin
3.4.1 Covering the MIP Using a Solid Phase Extraction Cartridge
3.4.2 Covering the MIP Using a Glass Tube
3.4.3 Procedure for Covering Magnetic MIPs
3.4.4 Procedure for Covering MIP Fiber
3.5 Protein Exclusion Tests
3.5.1 Column Packed with RAMIP
3.5.2 Solid Phase Extraction Cartridges
3.5.3 Solid Phase Microextraction RAMIP Fibers
3.6 Possible Applications
3.6.1 Solid Phase Extraction in Cartridge
3.6.2 Magnetic Dispersive Solid Phase Extraction
3.6.3 Microextraction by Packed Sorbents
3.6.4 Solid Phase Microextraction
3.6.5 In-Tube Solid Phase Microextraction
3.6.6 Column Switching Liquid Chromatography
4 Notes
References
Chapter 6: Synthesis of Double-Layer Imprinted Polymers: BSA Depletion
1 Introduction
2 Materials
2.1 Synthesis of BSA-Specific Double Imprinted Polymers (BSA-DLIPs)
2.2 Cleaning the Columns and Template Removal Studies
2.3 Characterization Studies
2.4 Adsorption Studies
2.5 Selectivity Studies
2.6 Desorption Studies
3 Methods
3.1 Synthesis of BSA-DLIPs
3.2 Cleaning the Columns and Template Removal Studies
3.3 Characterization Studies
3.4 Adsorption Studies
3.5 Selectivity Studies
3.6 Desorption Studies
4 Notes
References
Chapter 7: Magnetic MIPs: Synthesis and Applications
1 Introduction
2 Materials
2.1 Co-precipitation Method (Fe3O4) (See Note 1)
2.2 Silanization of Fe3O4
2.3 Functionalization of Fe3O4@SiO2
2.4 Computer Simulation to Choose the Functional Monomer and/or Solvent
2.5 Polymerization: Synthesis of MIP on Fe3O4@SiO2-C=C
2.6 Extraction of Template from M-MIP Particles
2.7 Sample Application
2.8 Electrochemical Measurements
3 Methods
3.1 Synthesis of Magnetic Nanoparticles
3.2 Molecular Modeling
3.3 Development and Characterization of MMIPs
3.4 Characterization
3.5 Application of MMIP as Dispersive Solid-Phase Extractor (DSPE) for the Determination of Ciprofloxacin in Bovine Milk
3.6 Application of MMIP Particles Previously Captured by a Magnetic Sensor for the Development of an Electrochemical Sensor
4 Notes
References
Chapter 8: Water-Compatible Fluorescent Molecularly Imprinted Polymers
1 Introduction
2 Materials
2.1 Synthesis of QD-Labeled Fluorescent MIP Nanoparticles with Surface-Bound Alkyl Halide Groups (i.e., ATRP-Initiating Groups...
2.2 Synthesis of QD-Labeled Fluorescent MIP Nanoparticles with Hydrophilic Poly(GMMA) (PGMMA) Brushes (i.e., QD-SiO2@MIP@PGMMA...
2.3 Synthesis of Organic Fluorophore-Labeled Fluorescent MIP nanoparticles with PHEMA Brushes (i.e., OF-MIP@PHEMA) via Hydroph...
3 Methods
3.1 The First Step of Two-step Approach´´: Synthesis of QD-SiO2@MIP (and Its Non-imprinted or Control Polymer (QD-SiO2@CP)) ... 3.2 The Second Step ofTwo-step Approach´´: Synthesis of QD-SiO2@MIP@PGMMA and QD-SiO2@CP@PGMMA via Surface-Initiated ATRP (...
3.3 ``One-step Approach´´: Synthesis of OF-MIP@PHEMA via Hydrophilic Macro-CTA-Mediated RAFTPP (See Notes 1, 3, and 5)
3.4 Calibration Measurements with Fluorescent MIP Nanoparticles with Hydrophilic Polymer Brushes (or Briefly Hydrophilic Fluor...
3.5 Direct and Highly Selective Optosensing of Tc in Complex Biological Samples with the Hydrophilic Fluorescent MIP Nanoparti...
4 Notes
References
Chapter 9: Generation of High-Affinity Aptamer-MIP Hybrid Nanoparticles
1 Introduction
2 Materials
2.1 Synthesis of Polymerizable Base
2.2 Synthesis of Modified Aptamer
2.3 Preparation of Template-Modified Solid-Phase Beads
2.4 Synthesis of AptaMIP NPs via the Template-Linked Solid-Phase Method
3 Methods
3.1 Synthesis of Base
3.2 Sequence Synthesis
3.3 Preparation of Protein-Modified Solid-Phase Beads
3.4 Preparation of Epitope-Modified Solid-Phase Beads
3.5 Synthesis of AptaMIP NPs
3.6 Selection of High-Affinity AptaMIP NPs
4 Notes
References
Chapter 10: Molecularly Imprinted Solid-Phase Extraction Sorbents for the Selective Extraction of Drugs from Human Urine
1 Introduction
2 Materials
2.1 Synthesis of MISPE Sorbent
2.2 Soxhlet Extraction
2.3 Batch Rebinding Studies
2.4 MISPE Protocol
2.5 Analysis of Human Urine
3 Methods
3.1 Synthesis of MISPE Sorbent
3.2 Batch Rebinding Experiments
3.3 MISPE Protocol
3.4 Analysis of Human Urine
3.5 Analytical Parameters
4 Notes
References
Chapter 11: Molecularly Imprinted Polymers Coupled with Surface-Enhanced Raman Spectroscopy to Detect Chemical Hazards in Foods
1 Introduction
2 Materials
2.1 Polymer Synthesis
2.2 Molecularly Imprinted Solid-Phase Extraction (MISPE)
2.3 Preparation of SERS Substrate
2.4 SERS Spectral Collection and Data Analysis
2.5 Detection of Chemical Hazard in Food Using MIP-SERS sensor
3 Methods
3.1 Synthesis of MIPs (and Non-imprinted Polymers [Optional])
3.2 Preparation of Molecularly Imprinted Solid-Phase Extraction (MISPE)
3.3 Synthesis of SERS Substrate
3.4 SERS Spectral Collection and Data Analysis
3.5 Detection of Pesticide in Food with Developed MIP-SERS Sensor (Fig. 1)
4 Notes
References
Chapter 12: Molecularly Imprinted Polymer for a Smart Dispersive Micro-Solid Phase Extraction Technique for Assessing Trace Le...
1 Introduction
2 Materials
2.1 MIP Synthesis
2.2 Template Removal from MIP
2.3 Characterization of MIP Beads
2.4 AFs Extraction from Culture Fish Flesh
2.5 Dispersive Micro-Solid Phase Extraction
2.6 Liquid Chromatography-Tandem Mass Spectrometry Measurements
3 Methods
3.1 MIP Synthesis and Characterization
3.2 AFs Extraction from Fish by Ultrasound-Assisted Extraction (UAE)
3.3 Dispersive Micro-Solid Extraction (D-μ-SPE)
3.4 Liquid Chromatography-Tandem Mass Spectroscopy (LC-MS/MS)
3.5 Optimization of the D-μ-SPE Parameters
3.6 Analytical Performance
4 Notes
References
Chapter 13: Preparation of Monolithic Fibers in Fused Silica Capillary Molds for Molecularly Imprinted Solid-Phase Microextrac...
1 Introduction
1.1 Overview
1.2 General Considerations in Monolithic Fiber Preparation
2 Materials
2.1 Polymerization Mixture
2.2 Preparation of Molecularly Imprinted Fibers
3 Methods
3.1 Preparation of the Fused-Silica Capillaries Used as Molds
3.2 Preparation of the Polymerization Mixture
3.3 Preparation of Molecularly Imprinted Fibers
4 Notes
References
Chapter 14: Quick, Easy, Cheap, and Effective Method for the Synthesis of Rugged Magnetic Molecularly Imprinted Stir-Bars
1 Introduction
2 Materials
2.1 Synthesis of Ferric Oxide (Fe3O4) Nanoparticles
2.2 Polymeric Mixture
2.3 Polymerization Equipment and Other Material
2.4 Template Removal, Conditioning, and Optimization of the (MMIP-SB)
3 Methods
3.1 Synthesis of Magnetite by Co-Precipitation
3.2 Preparation of Pre-Polymerization Mixture
3.3 Preparation of the Magnetic Molecularly Imprinted Stir-bar (MMIP-SB)
3.4 MMIP-SB Template Removal and Optimization
4 Notes
References
Chapter 15: Determination of Neopterin as a Prognostic Indicator Using Neopterin-Imprinted Cryogel Membranes
1 Introduction
2 Materials
2.1 Preparation of Neo-ImprintedCryogel Membranes (Neo-mip)
2.2 Cleaning the Cryogel Membranes and Template Removal Studies
2.3 Characterization of Neo-mip and NIP Cryogel Membranes
2.4 Adsorption Studies
2.5 Evaluation of Selective Adsorption Properties of Neo-mip for Neo
2.6 Desorption Studies
2.7 Recognition of Neo from Human Serum
3 Methods
3.1 Preparation of Neo-imprinted Cryogel Membranes (Neo-mip)
3.2 Cleaning the Cryogel Membranes and Template Removal Studies
3.3 Characterization of Neo-MIP and NIP Cryogel Membranes
3.4 Adsorption Studies
3.5 Evaluation of Selective Adsorption Properties of Neo-mip for Neo
3.6 Desorption Studies
3.7 Recognition of Neo from Human Serum
4 Notes
References
Chapter 16: Fluorescence Sensing with Molecularly Imprinted Polymer-Capped Quantum Dots
1 Introduction
2 Materials
2.1 Quantum Dots
2.2 Water Solubilization of the QDs
2.3 Molecularly Imprinted Polymer-Capped Quantum Dots (MIP@QDs)
2.4 Template Removal from MIP@QDs
3 Methods
3.1 Preparation of QDs
3.1.1 Preparation of Precursors
3.1.2 Synthesis of QDs
3.1.3 Water Solubilization of the QDs with Thiol Ligands
3.2 Synthesis of MIP@QDs
3.3 Optimization of Sensing Variables
3.3.1 Concentration of MIP@QDs
3.3.2 Effect of Incubation Time
3.4 Fluorescence Sensing of the Template Using MIP@QDs in a Standard Solution
3.5 Determination of the Template in Water Matrices Using Fluorescence Sensing of MIP@QDs
4 Notes
References
Chapter 17: Dual-Fluorescent Nanoparticle Probes Consisting of a Carbon Nanodot Core and a Molecularly Imprinted Polymer Shell
1 Introduction
2 Materials
2.1 Synthesis of Red-Emissive Carbon Nanodots (R-CNDs)
2.2 Synthesis of Fluorescent Monomer (I)
2.3 Synthesis of Red Carbon Nanodot-Doped Silica Nanoparticles (R-CSNs)
2.4 RAFT Agent Functionalization
2.5 MIP Synthesis
2.6 Equipment
2.7 Characterization and Evaluation of MIP Binding Performance (Recommended)
3 Methods (See Note 1)
3.1 Synthesis of Fluorescent Probe Monomer (I) (See Note 1)
3.2 Synthesis of R-CNDs
3.3 Synthesis of R-CSNs
3.4 RAFT Agent Functionalization
3.4.1 APTES Functionalization
3.4.2 4-Cyano-4-(Phenylcarbonothioylthio)Pentanoic Acid (CPDB) Functionalization
3.5 MIP Synthesis (See Note 6)
3.6 Characterization and Evaluation of CGA-MIP Binding Performance (Recommended)
3.6.1 Size Determination (See Note 6)
3.6.2 Zeta Potential Determination (See Note 8)
3.6.3 APTES and CPDB Determination
3.6.4 Evaluation of MIP Binding Performance by Fluorescence Rebinding Test (Fig. 3)
4 Notes
References
Chapter 18: Molecularly Imprinted Polymer-Based Quartz Crystal Microbalance Sensor for the Clinical Detection of Insulin
1 Introduction
2 Materials
2.1 Reagents
2.2 Equipment
2.3 Solutions
3 Methods
3.1 Thiol Groups Modification of QCM Chip
3.2 Preparation of QCM Sensor
3.3 Characterization Studies
3.4 Kinetic Analysis
3.5 Isotherm Models
3.6 Detection of Insulin from Artificial Plasma
3.7 Selectivity
3.8 Reusability
4 Notes
References
Chapter 19: Preparing Selective Nanozymes by Molecular Imprinting
1 Introduction
1.1 Nanozymes
1.2 Applications of Nanozymes
1.3 The Specificity Problem in Nanozymes
1.4 Increasing Nanozyme Specificity by Molecular Imprinting
2 Materials
2.1 Preparation of Iron Oxide Nanoparticles (Fe3O4 NPs)
2.2 Preparation of MIP-Coated Iron Oxide Nanoparticles (MIP-Fe3O4 NPs)
2.3 Template Removal
2.4 Preparation of Substrate Solutions
2.5 Activity Assays
2.6 Selectivity Tests
3 Methods
3.1 Preparation of Iron Oxide Nanoparticles
3.2 Preparation of MIP-Coated Iron Oxide Nanoparticles (MIP-Fe3O4 NPs)
3.3 Template Removal
3.4 Activity Assays
3.5 Selectivity Tests
4 Notes
References
Chapter 20: Enzyme-Mimics Molecularly Imprinted Polymers Based on Metal Complexes: Electropolymerization and Electrocatalytic ...
1 Introduction
2 Materials
2.1 Electrode Preparation
2.2 Electropolymerization
2.3 Washing for Template Removal
2.4 Electrochemical (Voltammetric and Amperometric) Measurements
3 Methods
3.1 MIP and NIP Electropolymerization
3.2 Calibration Measurements with Voltammetric MIP-Sensor
3.3 Calibration Measurements with Amperometric MIP-Sensor
4 Notes
References
Chapter 21: Using Molecular Dynamics in the Study of Molecularly Imprinted Polymers
1 Introduction
2 Materials
3 Methods
3.1 System Design
3.2 Input Generation
3.2.1 Virtual Molecular Models
3.2.2 Generating GAFF Parameters
3.2.3 Constructing the Virtual Simulation System
3.2.4 Applying Parameters
3.3 Simulating Constructed Systems
3.3.1 Energy Minimization
3.3.2 Temperature Equilibration
3.3.3 Pressure Equilibration
3.3.4 Data Production
3.4 Processing and Analysis of Data
3.4.1 Minimization and Equilibration Data Analysis
3.4.2 Production Data Analysis-Cpptraj
4 Notes
References
Chapter 22: The Search for Peptide Epitopes for Molecular Imprinting Through Bioinformatics
1 Introduction
2 Materials
3 Methods
3.1 Selection of Immunogenic Peptide Epitopes
3.2 Rational Selection of Linear Peptide Epitopes
3.3 Selection of Structured Epitopes
4 Conclusions
References
Index