Published by
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Handbook of Molecular Imprinting: Advanced Sensor Applications
Copyright © 2013 by Pan Stanford Publishing Pte. Ltd.
All rights reserved. This book, or parts thereof, may not be reproduced in any formor by any means, electronic or mechanical, including photocopying, recording
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For photocopying of material in this volume, please pay a copying fee throughthe Copyright Clearance Center, Inc., 222 Rosewood Drive, Danvers, MA 01923,
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ISBN 978-981-4316-65-1 (Hardcover)ISBN 978-981-4364-32-4 (eBook)
Printed in the USA

Contents

Part 1

Fundamentals of Molecular Imprinting and
Sensor Applications

1. Fundamentals and Perspectives of Molecular
Imprinting in Sensor Applications
		 Seung-Woo Lee, Sergiy Korposh, Roman Selyanchyn, andToyoki Kunitake
1.1 Introduction
1.2 Molecular Imprinting in Organic Matrices
1.2.1 Covalent Approach
1.2.2 Non-Covalent Approach
1.2.3 Other Approaches for Organic
MIP Fabrication
1.3 Molecular Imprinting in Inorganic Matrices
1.3.1 Sol-Gel Approach
1.3.2 Liquid Phase Deposition (LPD) Approach
1.4 Major Transducers
1.4.1 Mass-Sensitive Transducer
1.4.2 Electrochemical Transducer
1.4.3 Optical Transducer
1.5 Applications of Organic MIP Materials in Sensors
1.6 Applications of Inorganic MIP Materials in Sensors
1.6.1 Silica Based Imprinted Materials
1.6.2 Hybrid Imprinted Materials
1.6.3 LPD Based Imprinted Materials
1.6.4 Strategy for Improved Selectivity
1.7 Conclusions

2. Molecularly Imprinted Optical Sensing Receptor
		 Sing Muk Ng and Ramaier Narayanaswamy
2.1 Introduction
2.2 Chronological Protocols and Procedures

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2.2.1 Selection of Materials and Ingredients
2.2.2 Polymerization Options
2.2.2.1 Free-radical initiatedpolymerization
2.2.2.2 Condensation polymerization
2.2.2.3 Electropolymerization
2.2.3 Configurations of Matrix
2.2.3.1 Bulk
2.2.3.2 Monoliths
2.2.3.3 Membranes
2.2.4 Handling and Preparation
2.3 Rational Design of Receptors
2.3.1 Interaction Study of Pre-Polymerization
Ingredients
2.3.2 Computational Modeling2.3.3 Thermodynamic Considerations
2.3.4 Repeatability and Reproducibility
2.3.5 Commercialization
2.4 Optical Sensing Schemes and Transduction
Systems
2.4.1 Direct Monitoring of Analyte
2.4.2 Direct Fluorescence
2.4.3 Displacement Assay
2.4.4 Reflectance and Absorbance
2.4.5 Phosphorescence
2.4.6 Chemiluminescence
2.4.7 Surface Plasmon Resonance2.4.8 Fluorescence Lifetime Decay
2.5 Advanced Probe Designs and Sensing
Configuration
2.5.1 Sensor Arrays
2.5.2 Optical MIP Chips
2.5.3 Micro- and Nano-sized Sensors
2.6 Binding Aspects and Analytical Signals
2.6.1 Binding Isotherms and Affinity
Distributions
2.6.2 Batch Binding Analysis and Binding
Models

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2.6.3 Correlation of Analytical Signal with
Binding Isotherms Models
2.6.4 Advantage and Limitation
2.7 Summary

3. Translational Applications of Molecularly
Imprinted Polymer-Based Electrochemical
Sensors
		 Hung-Yin Lin, James L. Thomas, and Mei-Hwa Lee
3.1 Introduction
3.2 Principle of Molecularly Imprinted Polymers
3.2.1 Synthesis of MIPs
3.2.2 Characterization of MIPs
3.2.3 Morphology of MIPs
3.3 Transducers Employed with Molecularly
Imprinted Polymers as Sensing Elements
3.3.1 Types of Transducers
3.3.2 Interface of Transducer and Molecularly
Imprinted Polymers
3.3.3 Miniature MIPs-Based Sensors
3.3.4 Demonstration of MIPs-BasedElectrochemical Sensors
3.4 Molecularly Imprinted Polymers-Based Sensors
for the Real World
3.4.1 Source of Real Samples
3.4.2 Biomarkers
3.4.3 Cross-Talk Interference
3.5 Prospective

4. Optical Sensors for MonitoringTrace Inorganic
Toxins		 T. Prasada Rao, Dhanya James, and Milja T. Elias
4.1 Environmental Trace Analysis
4.2 Inorganic Toxins
4.3 Importance of Sampling in Trace Analysis
4.3.1 Sample Handling
4.3.2 Sample Pre-Treatment, Homogenizationand Sub-Sampling
4.3.3 Sample Preparation

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4.3.3.1 Decomposition of inorganic ororganic matrices
4.3.3.2 Separation and pre-concentration
steps
4.4 Trace/Ultra Trace Analytical Techniques
4.4.1 Selection of Analytical Technique/Method
4.4.2 Essential Features of Analytical
Techniques
4.4.2.1 Signal processing, data handlingand reporting
4.4.2.2 Signal integrity
4.4.2.3 Data handling
4.4.2.4 Good Automated Laboratory
Practice (GALP) [1]
4.4.2.5 Reporting of results
4.5 Chemical Speciation
4.6 Sensors
4.6.1 Fundamentals of Optical Sensors(Optodes)4.6.2 Optical Sensing of Ionic Analytes
4.6.3 Optical Sensing of Neutral Analytes
4.7 Molecularly Imprinted Polymers
4.7.1 Molecular Imprinting Technology
4.7.2 MIPs in Optical Sensing
4.8 Optical Sensors vis-a-vis Other Sensor Techniques
4.9 Future Outlook

5. MIP Thermistor
		 Rajagopal Rajkumar, Umporn Athikomrattanakul,
Kristian Lettau, Martin Katterle, Bengt Danielsson,
Axel Warsinke, Nenad Gajovic-Eichelmann, andFrieder W. Scheller
5.1 Introduction
5.1.1 The MIP Concept
5.1.2 MIP Sensors
5.1.3 Enzyme Thermistors
5.2 Covalently Imprinted Polymers Using Boronic
Acid Derivates5.2.1 Synthesis of Template (Fructosyl Valine)

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5.2.2 Synthesis of Functional Monomer (Vinyl
Phenyl Boroxine)
5.2.3 Synthesis of N-[β-D-Fructopyranosyl-(1)]L-Valine2,3; 4,5-bis-O- ((4-Vinylphenyl)
Boronate)
5.2.4 Synthesis of MIP and Control Polymers
5.2.5 MIP Thermistor Set-Up and Measurements
5.2.6 Thermometric MIP Sensor for Fructose
5.2.7 Thermometric MIP Sensor for Fructosyl
Valine5.2.8 Concentration Dependence of Fru-Val
Binding
5.2.9 Closed Loop Studies
5.3 Non-Covalent MIPs Containing Two Functional
Monomers for Carboxyphenyl Aminohydantoin(CPAH) as Analogon of Nitrofurantoin (NFT)
5.3.1 Synthesis of an Analogue Template,
Carboxyphenyl Aminohydantoin (CPAH)
5.3.2 Synthesis of Functional Monomers
5.3.3 Preparation of MIPs Based on Two
Functional Monomers
5.3.4 MIP-Based Thermometric Study
5.4 Bi-Functional Esterolytically Active MIP
5.4.1 Polymer Preparation
5.4.2 Thermometric Characterization ofAdsorption and Catalysis
5.5 Conclusions

Part 2

Potential Materials for Molecular
Imprinting

6. The Use of a Thermally Reversible Bond for
Molecular Imprinting
		 Ji Young Chang
6.1 Introduction
6.2 Cross-Linked Vinyl Polymer Matrix
6.3 Silica Matrix
6.4 Polyimide as Noncross-Linked Matrix
6.5 Summary and Outlook

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7. Molecular-Sieving Silica/Tin Oxide Sensor
Prepared by Chemical Vapor Deposition in the
Presence of Template Molecule
		 Naonobu Katada and Miki Niwa
7.1 Introduction
7.2 Preparation Method
7.3 Selective Chemisorption
7.4 Sensing Function
7.4.1 Selectivity for Various Molecules
7.4.2 Improved Response and Selectivity ofFilm-Type Sensor
7.4.3 Detection of Dialkyl Phthalate
7.5 Conclusion

8. Environmental Approaches by Molecular
Imprinting on Titanium Dioxide
		 Milka Nussbaum and Yaron Paz
8.1 Introduction
8.2 Preparation
8.2.1 Preparation of Molecularly Imprinted (MI)Structures
8.2.1.1 Sol-gel method using alkoxidechemistry
8.2.1.2 “Layer-by-layer” approach using
sol-gel method
8.2.1.3 Liquid phase deposition/chemicalbath deposition
8.2.1.4 Titanyl sulfate chemistry
8.2.1.5 Titanium tetrachloride chloride
chemistry
8.2.1.6 Potassium titanyl oxalatechemistry
8.2.2 Preparation of Molecularly Imprinted
Polymer (MIP) Structures
8.2.3 Preparation of Molecularly Imprinted
Inorganic (MII) Structures
8.2.4 Preparation of Molecularly Imprinted
Host (MIH) Structures
8.3 Characterization

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8.3.1 Interaction Between Host and Template
8.3.2 Assessment of Removal of Template
Molecules
8.3.3 Concentration and Structure of Sites
8.3.4 Surface Area
8.3.5 Film Morphology and Particles’ Size
8.3.6 Film Thickness
8.3.7 Crystalline Phase
8.4 Applications
8.4.1 Sensing
8.4.1.1 Quartz crystal microbalance(QCM) sensors
8.4.1.2 Electrochemical sensors
8.4.1.3 Ion sensitive field effecttransistors
8.4.1.4 Matrix-assisted laser desorption/
ionization (MALDI)massspectrometry
8.4.1.5 Evaluation of performance ofsensors
8.4.1.5.1 Sensitivity
8.4.1.5.2 Imprinting efficiency
ratio
8.4.1.5.3 Selectivity
8.4.1.5.4 Response time
8.4.1.5.5 Repeatability
8.4.1.5.6 Stability andreproducibility
8.4.2 Photocatalysis
8.4.2.1 Introduction
8.4.2.2 Performance
8.4.2.2.1 Efficiency
8.4.2.2.2 Selectivity
8.4.2.2.3 Adsorption vs.
photocatalytic
degradation
8.4.2.2.4 Imprinting ofpseudo-target molecules
8.4.2.2.5 Intermediates andby-products

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8.4.2.2.6 Stability andrepeatability
8.4.3 Separation by Selective Filtration
8.5 Conclusion

Part 3

MIP Sensors for Environmental
and Trace Detection

9. Molecularly Imprinted Nanocomposites for Highly
Sensitive SPR Detection		 Jun Matsui and Kensuke Akamatsu
9.1 Introduction
9.2 Surface Plasmon Resonance of Metal Nanoparticles
9.2.1 Biosensors Utilizing Gold Nanoparticles
9.2.2 Synthesis of Surface-Functionalized Gold
Nanoparticles
9.3 Molecularly Imprinted Nanocomposite
9.3.1 Concept
9.3.2 Colorimetric Sensing with Molecularly
Imprinted Nanocomposite
9.3.2.1 Physical properties
9.3.2.2 Selectivity
9.3.3 SPR Sensor with Molecularly Imprinted
Nanocomposite Gel
9.3.3.1 Preparation of sensor chip
9.3.3.2 SPR measurement
9.3.4 SPR Sensing of Atrazine
9.3.4.1 Preparation of sensor chip
9.3.4.2 Effects of gold nanoparticles on
sensitivity
9.3.4.3 Effects of molecular imprinting
on sensitivity
9.3.4.4 Selectivity
9.4 Conclusion

10. Molecularly Imprinted Room Temperature
Phosphorescent Optosensors for Environmental
Pollutants
		 He-Fang Wang and Xiu-Ping Yan
10.1 Introduction

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10.2 Fabrication of MIPs-Based RTP Optosensors
10.2.1 Conventional MIPs with RTP Detection
10.2.2 Incorporation of Heavy Atoms into MIPs
10.2.3 Surface Imprinting on Phosphorescent
Nanoparticles
10.3 RTP Detection Modes
10.3.1 Static Measurement
10.3.2 Flow-Through Assays
10.4 Applications
10.4.1 RTP Sensing of Nafcillin
10.4.2 RTP Sensing of PAHs
10.4.3 Sensing of Copper Ions
10.4.4 Sensing of Pentachlorophenol
10.5 Conclusions

11. Electrochemical Sensing of Nitroaromatic
Compounds in Natural Waters and Soil Samples
		 Taher Alizadeh
11.1 Introduction
11.2 Nitroaromatic Compounds
11.3 Different Strategies Developed for Preparationof MIP for Nitroaromatic Compounds
11.4 Electrochemistry of Nitroaromatic Compounds
11.5 MIP-Based Electrochemical Sensors
11.5.1 Polymerization Directly on the ElectrodeSurface11.5.1.1 Sol-gel and poly TiO2 systems
11.5.1.2 Electrodes modified withelectropolymerized films
11.5.2 Coupling of the MIP Particles with
Electrochemical Transducers
11.5.3 MIP as a Solid Phase Sorbent for
Separation and Preconcentration
Before Electrochemical Determination

12. Trace Detection Based on Cyclodextrin Anchored
Molecularly Imprinted TiO2 Thin Films
		 Seung-Woo Lee
12.1 Introduction
12.2 Surface Sol-Gel Process and Molecular Imprinting

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12.3 Two-Dimensional Imprinting with Juxtaposed
CD Hosts on Metal Oxide Surface
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12.3.1 BPA Sensing Based on Electrochemical
Impedance Changes
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12.3.2 SAM Effects on cSPI Response429
12.3.3 cSPI Response to BPA on a BPA-Imprinted
TiO2/β-CD Film
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12.3.4 Selectivity of the BPA-Imprinted
TiO2/β-CD Film
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12.4 Trace Detection of Explosives Using a CD-Anchored
Metal Oxide Matrix
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12.4.1 Surface Modification for 2,4-DNT
Recognition436
12.4.2 cSPI Response and Sensitivity to 2,4-DNT
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12.5 Imprinting Process Associated with a Gas Phase
Sol-Gel Technique439
12.5.1 Gas-Phase Surface Sol-Gel (GSSG) Process 439
12.5.2 Three-Dimensional Assembly of
Templated CDs with TiO2 Ultrathin Layers 440
12.5.3 BPA Response and Imprinting Effect441
12.6 Future Prospects
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13. Molecularly Imprinted Au Nanoparticle Composites
and Their Application for Sensing, Controlled
Release, and Photoelectrochemistry
		 Itamar Willner and Ran Tel-Vered
13.1 Introduction
13.2 Imprinting of Molecular Recognition Sites in Au
NP Composites via Donor–Acceptor and/orElectrostatic Interactions
13.3 Imprinted Ligand-Functionalized bis-AnilineCrosslinked Au NP Composites for Sensing
13.4 Electrochemically Triggered Imprinted Au NP“Sponges”
13.5 Controlling the Wettability of Surfaces by
Imprinted Au NP Composites
13.6 Imprinted Semiconductor Nanoparticle/
Metal Nanoparticle Composites for Enhanced
Photoelectrochemistry

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13.7 Conclusions and Perspectives

Part 4

Bio-Medical Applications of MIP Sensors

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14. Protein-Sensing Using Organic/Inorganic Hybrid
Materials Prepared by Liquid-Phase DepositionBased Molecular Imprinting
487
		 Toshifumi Takeuchi
14.1 Introduction
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14.2 Liquid-Phase Deposition489
14.3 Cationic Polyions/Titanium Oxide Hybrid Materials
for Acidic Protein Recognition490
14.4 Anionic Polymer/TiO2 Hybrid Materials for Basic
Protein-Sensing
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14.5 Protein Imprinting on Quantum Dots by LPD
Method493
14.6 Conclusion
494
15. Molecular Imprinted Polymer-Based
Chemiluminescence Sensors
		 Zhujun Zhang
15.1 Introduction
15.2 Molecular Imprinted Polymer-Based
Chemiluminescence Flow Sensors
15.3 Molecular Imprinted Polymer-BasedCL Imaging
Sensors
15.4 Molecular Imprinted Polymer-Based
Chemiluminescence Biosensor Chips
15.5 Molecular Imprinted Polymer Recognition and
Online Electrogenerated Reagent
Chemiluminescence Detection

16. Detection of Cells and Viruses Using Synthetic
Antibodies
		 Adnan Mujahid, and Franz L. Dickert
16.1 Introduction
16.2 Molecular Imprinting for Biomolecules
16.3 Synthetic Antibodies for Cells Detection
16.3.1 Yeast Cells

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16.3.2 Blood Cells Detection
16.3.3 Bacterial Detection Through Synthetic
Materials
16.4 Imprinted Materials – A novel tool for Virus
Sensing
16.4.1 Tobacco Mosaic Virus (TMV) Imprinting
16.4.2 Parapox Ovis Virus (ORF) Sensing
16.4.3 Bovine Leukemia Virus Imprinting
16.4.4 Molecular Imprinting for Picornaviruses
16.4.5 Dengue Virus Detection Through Epitope
Imprinting Approach
16.4.6 Poliovirus Detection by Imprinted SAMs
16.4.7 Virus Sensing by MIP-CoatedMicrosensors
16.5 Concluding Remarks and Future Outlook

Part 5

MIP Market and Prospects

17. Molecularly Imprinted Polymers: Science Goes
Market? A Market Analysis Based on the Patent
Situation
		 Soeren Schumacher
17.1 Introduction
17.2 Methods
17.3 Results and Discussion
17.3.1 Patent Activity
17.3.2 Application Areas
17.3.3 Citation Analysis
17.3.4 S-Curve Consideration
17.3.5 Company Overview
17.3.6 Patent Portfolio of Companies
17.3.7 Competitive and SWOT Analysis
17.4 Conclusion
Index			

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PrefaceMolecular imprinting is now established as an indispensable tool
for separation and sensor technologies. The most popular scheme
to realize the molecular imprinting concept is probably the crosslinking of linear polymers in the presence of template molecules.
Three-dimensional cavities would be created in the polymer
network after the removal of template molecules. Unfortunately,
the conformational adaptability of linear polymers is limited, andtemplate molecules and the surrounding polymer chains cannot
produce best molecular fitting in most cases. Molecular fitting bycommonly employed imprinting materials is less than satisfactorycompared with polypeptide chains of protein molecules. Superior
three-dimensional fit of active sites of enzyme molecules and their
specific substrates are truly remarkable.
Some of the inorganic chains are much more flexible than organic
chains. For example, rotational and bending motions of metal–oxygenbonds are more facile than those of carbon–carbon bonds. This
fact implies that metal oxide gels provide better molecular fittingwith template molecules. In fact, the first example of molecularimprinting was reported in 1931 for silica matrix, as mentioned in
Chapter 1. The use of inorganic matrices became much expanded
in recent years and now occupies a significant fraction of molecularimprinting studies.
This handbook reflects this situation and attempts to survey the
recent advances of molecular imprinting in inorganic and organic
matrices in a combined form. The functional aspect is more or lessfocused on sensor applications. Such applications have become
increasingly important in relation to environmental and biomedical
issues, as summarized as Parts 3 and 4, respectively. The discussionin Chapter 1 will help grasp the pros and cons of inorganic matrices
relative to organic matrices, and Part 5 provides a patent-based
market analysis of molecular imprinting technology.
We are grateful to Mr. Stanford Chong of Pan Stanford Publishingfor his continuous encouragement during the preparation of thishandbook. Many thanks also to our wives, who showed splendid
perseverance towards the absent (minded) husbands.

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