Enzyme-linked Immunosorbent Assay (ELISA), 1st ed. 2018
From A to Z

SpringerBriefs in Forensic and Medical Bioinformatics Series

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Language: English
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Support: Print on demand
This book offers comprehensive information on all aspects of ELISA, starting with the fundamentals of the immune system. It also reviews the history of analytical assays prior to the advent of ELISA (enzyme-linked immunosorbent assay) and addresses the materials of choice for the fabrication of the platforms, possible biomolecular interactions, different protocols, and evaluation parameters. The book guides readers through the respective steps of the analytical assay, while also familiarizing them with the possible sources of error in the assay. It offers detailed insights into the immobilization techniques used for protein attachment, as well as methods for evaluating the assay and calculating the key parameters, such as sensitivity, specificity, accuracy and limit of detection. In addition, the book explores the advantages and shortcomings of the conventional ELISA, as well as various approaches to improving its performance. In this regard, merging and integrating other technologies with widely known ELISAs have opened new avenues for the advancement of this immunoassay. Accordingly, the book provides cutting-edge information on integrated platforms such as ELISpot, plasmonic ELISAs, sphere-/bead-based ELISAs, paper-/fiber-based ELISAs and ELISA in micro-devices. 

1 Fundamentals of ELISA: from the evolution of the immunoassays until invention of ELISA
Abstract
1. 1 Evolution of the immunoassays until invention of ELISA 
1.1.1 Side chain theory
1.1.2 Antigen-antibody binding theory
1.1.3 Discovery of antibody structure
1.1.4 Invention of RIA
1.1.5 Invention and development of ELISA
1.2 Principles of the immune system
1.2.1 Antibody production in human body
1.2.2 Categories of antibodies
1.2.3 Antigen-antibody coupling
1.2.4 Specificity of the antigen-antibody coupling
1.3 Biomolecular interactions between antibody and antigen
1.3.1 Hydrogen bonding
1.3.2 Hydrophobic interaction
1.3.3 Ionic attraction
1.3.4 Van der Waals forces
1.3.4.1 London dispersion
1.3.4.2 Dipole-dipole interaction
1.3.4.3 Ion-dipole force
References 

2 General overview on applications of ELISA
Abstract
2.1 Application of ELISA
2.1.1 Food industry
2.1.2 Vaccine development
2.1.3 Immunology
2.1.3.1 Autoimmunity
2.1.3.2 Humoral immunity
2.1.4 Diagnosis
2.1.4.1 Home pregnancy test
2.1.4.2 Cancer detection
2.1.4.3 Detection of the infectious diseases
2.1.5 Toxicology 
2.1.6 Drug monitoring and pharmaceutical industry
2.1.7 Transplantation
References 

3 Step by step with ELISA: mechanism of operation, crucial elements, different protocols, and insights on immobilization and detection of various biomolecular entities 
Abstract
3.1 Mechanism of operation
3.2 Different elements of the assay
3.2.1 Solid phase
3.2.1.1 Adsorption
3.2.2 Washing process
3.2.3 Antigens 
3.2.4 Antibodies
3.2.4.1 Antispecies antibodies
3.2.5 Enzyme
3.2.5.2 Different types of enzyme
3.2.5.2 Enzyme conjugation
3.2.6 Substrate
3.2.7 Stopping process
3.2.8 Reading process
3.2.8.1 Chromogenic assay
3.2.8.2 Chemifluorescenct assay
3.2.8.3 Chemiluminescent assay
3.2.8.4 Reading apparatus
3.2.9 Controls
3.3 Target biomolecular entities by ELISA
3.4 Different protocols
3.4.1 Direct ELISA
3.4.2 Indirect ELISA
3.4.3 Sandwich ELISA
3.4.4 Double sandwich ELISA
3.4.5 Competitive ELISA
3.5 Immobilization techniques for protein attachment 
3.5.1 Physical immobilization
3.5.2 Covalent immobilization
3.5.2.1 Immobilization via zero-Length cross linker
3.5.2.2 Immobilization via spacers
3.5.3 Immobilization via entrapment
3.5.4 Oriented immobilization
References 

4 Evaluation of the detection results obtained from ELISA
Abstract
4.1 Conducting a reliable assay
4.1.1 Sources of error
4.1.2 Troubleshooting 
4.2 Key parameters for evaluation of the assay
4.2.1 Sensitivity
4.2.2 Specificity
4.2.3 Accuracy
4.2.4 Limit of detection
4.3 Measurable units in ELISA
References 

5 Advantages, disadvantages and modifications of conventional ELISA 
Abstract
5.1 Significance of conventional ELISA 
5.2 Shortages of conventional ELISA
5.3 Materials of choice for fabrication of ELISA well plates
5.4 Different types of ELISA well plates
5.5 Modified ELISA platforms
5.5.1 ELISpot
5.5.2 Plasmonic ELISA
5.5.3 Sphere-/bead-based ELISAs
5.5.4 Paper-/fiber-based ELISAs
5.5.5 ELISA in microdevices
References 

6 Common questions and answers about ELISA 
Abstract
References 
Summary

Dr. Samira Hosseini obtained her B.Sc. degree in Applied Physics, her M.Sc. degree in Polymer Chemistry and her PhD degree in Biomedical Engineering. Currently, she is working as a Postdoctoral Researcher at the School of Engineering and Sciences, Tecnológico de Monterrey, Mexico. She is the author/co-author of more than 15 ISI indexed scientific publications, 12 book chapters and the inventor/co-inventor of 4 Intellectual properties. She is a member of the Mexican National System of Researchers. Her main areas of research are surface engineering, interface science, bioanalytical assays and polymeric materials. 

Dr. Patricia Vázquez-Villegas obtained her Bachelor’s degree in Food Sciences Engineering program in 2008 and her PhD in Engineering Sciences from Tecnológico de Monterrey, Monterrey, México in 2013 majoring in Biotechnology. She made a one-year internship in 2012 at the University of British Columbia, Vancouver, Canada under the supervision of Dr. Charles A. Haynes. Since 2014 she holds a Postdoc position in Tecnológico de Monterrey where she is working in the development and application of platforms for purification and detection of different kinds of biomarkers. Dr. Vázquez is the author/co-author of 8 ISI indexed scientific publications, 1 book chapter and the inventor/co-inventor of 4 intellectual properties. She is a member of the Mexican National System of Researchers. Her main contribution had been in the field of continuous recovery of biomolecules using liquid–liquid extraction.

Professor Rito-Palomares obtained his PhD in Chemical Engineering from The University of Birmingham, United Kingdom in 1995. Following a sabbatical in the Department of Chemical Engineering at the University of Cambridge, United Kingdom (2001)., Prof. Rito-Palomares is currently the Director of the Biotechnology Centre and a Full Professor at Tecnologico de Monterrey, Mexico. He is a member of the Mexican Academy of Sciences, scientific committee of the

Includes supplementary material: sn.pub/extras