Enzyme-Linked Immunosorbent Assay (Elisa): From A to Z

ISBN-13: 9789811067655 / Angielski / Miękka / 2018 / 115 str.

Samira Hosseini; Patricia Vazquez-Villegas; Marco Rito-Palomares

Opis

This book offers comprehensive information on all aspects of ELISA, starting with the fundamentals of the immune system. 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.

Spis treści

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

O autorze

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 International Foundation for Science (IFS, Sweden) as well as Mexican Research Council. He is the author/co-author of more than 100 ISI indexed scientific publications, 4 book chapters and inventor/co-inventor of 5 Intellectual properties that present his research work in the Bio-separation field. His main research contributions have been in the areas of biochemical recovery of biological products, and production, characterization and recovery of chemically modified proteins and the potential purification of stem cells. 

Sergio O. Martinez is Professor of the Electrical and Computer Engineering Department at Tecnológico de Monterrey, Monterrey Campus; and researcher and chair of the Sensors and Devices area of the National School of Engineering and Sciences at the same Institute. Sergio O. Martínez received the BS degree in Electronics and Communications Engineering and the MS degree in Control Engineering both from Tecnológico de Monterrey, Campus Monterrey, Mexico in 1983 and 1985, respectively. He obtained a MS degree in Electronics from Philips International Institute, Eindhoven, Netherlands in 1990; and the PhD degree in Microelectronics from National Polytechnic Institute of Grenoble, France in 2002. Prof. Martínez-Chapa is a member of the Mexican National System of Researchers since 2004 as well as member of the Mexican Academy of Sciences since 2010. His recent research activities, focused on Micro/Nanotechnologies, include Microfabrication Processes for Particle Manipulation Devices, Electrokinetically driven Microdevices for Manipulation of Suspended Particles in Fluidic Media, and Modeling and Design of surface plasmon resonance (SPR)-based Biosensing Platforms. He has authored/co-authored 75 peer-reviewed publications, 7 book chapters and the inventor/co-inventor of 2 granted intellectual properties.