Enzyme-linked Immunosorbent Assay (ELISA): From A to Z

124 Pages·2018·3.7 MB·English

by  Samira Hosseini

SPRINGER BRIEFS IN APPLIED SCIENCES AND
TECHNOLOGY  FORENSIC AND MEDICAL BIOINFORMATICS
Samira Hosseini
Patricia Vázquez-Villegas
Marco Rito-Palomares
Sergio O. Martinez-Chapa
Enzyme-linked
Immunosorbent
Assay (ELISA)
From A to Z
SpringerBriefs in Applied Sciences
and Technology
Forensic and Medical Bioinformatics
Series editors
Amit Kumar, Hyderabad, India
Allam Appa Rao, Hyderabad, India
More information about this series at http://www.springer.com/series/11910
á
Samira Hosseini Patricia V zquez-Villegas
(cid:129)
Marco Rito-Palomares Sergio O. Martinez-Chapa
(cid:129)
Enzyme-linked
Immunosorbent Assay
(ELISA)
From A to Z
123
Samira Hosseini MarcoRito-Palomares
Escuela deIngenieríay Ciencias Escuela deMedicina yCiencias deSalud
Tecnologico deMonterrey Tecnologico deMonterrey
Monterrey, NL Monterrey, NL
Mexico Mexico
Patricia Vázquez-Villegas SergioO.Martinez-Chapa
Escuela deIngenieríay Ciencias Escuela deIngenieríay Ciencias
Tecnologico deMonterrey Tecnologico deMonterrey
Monterrey, NL Monterrey, NL
Mexico Mexico
ISSN 2191-530X ISSN 2191-5318 (electronic)
SpringerBriefs inApplied SciencesandTechnology
ISSN 2196-8845 ISSN 2196-8853 (electronic)
SpringerBriefs inForensic andMedical Bioinformatics
ISBN978-981-10-6765-5 ISBN978-981-10-6766-2 (eBook)
https://doi.org/10.1007/978-981-10-6766-2
LibraryofCongressControlNumber:2017962065
©TheAuthor(s)2018
Thisworkissubjecttocopyright.AllrightsarereservedbythePublisher,whetherthewholeorpart
of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations,
recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission
orinformationstorageandretrieval,electronicadaptation,computersoftware,orbysimilarordissimilar
methodologynowknownorhereafterdeveloped.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this
publicationdoesnotimply,evenintheabsenceofaspecificstatement,thatsuchnamesareexemptfrom
therelevantprotectivelawsandregulationsandthereforefreeforgeneraluse.
The publisher, the authors and the editors are safe to assume that the advice and information in this
book are believed to be true and accurate at the date of publication. Neither the publisher nor the
authorsortheeditorsgiveawarranty,expressorimplied,withrespecttothematerialcontainedhereinor
for any errors or omissions that may have been made. The publisher remains neutral with regard to
jurisdictionalclaimsinpublishedmapsandinstitutionalaffiliations.
Printedonacid-freepaper
ThisSpringerimprintispublishedbySpringerNature
TheregisteredcompanyisSpringerNatureSingaporePteLtd.
Theregisteredcompanyaddressis:152BeachRoad,#21-01/04GatewayEast,Singapore189721,Singapore
Preface
ELISA:FromAtoZwaswrittenaimingtoprovideReaderswithapropercoverage
ofallaspectsofELISAfromfundamentalsofimmunesystemandthehistoryofthe
analytical assay prior to the invention of ELISA to the materials of choice for
fabricationoftheplatforms,possiblebiomolecularinteractions,different protocols,
and evaluation parameters among the rest. This book guides Readers through dif-
ferent steps of the analytical assay while familiarizing them with the possible
sources of error in the assay. The book offers detailed insights on the immobi-
lization techniques used for protein attachment, different methods for evaluation
oftheassayandcalculationofthekeyimportantparametersintheanalyticalassay
such as sensitivity, specificity, accuracy, and limit of detection. Advantages and
shortagesoftheconventionalELISAaswellasdifferentattemptsforimprovement
of its performance are covered in this book. Merging and intergrading different
technologies with widely known ELISA have opened numerous windows of
opportunity to the advancement of this immunoassay. In that respect, the current
book provides the latest updates on integrated platforms such as ELISpot, plas-
monic ELISA, sphere-/bead-based ELISAs, paper-/fiber-based ELISAs as well as
ELISA in micro-devices.
Monterrey, Mexico Samira Hosseini
Patricia Vázquez-Villegas
Marco Rito-Palomares
Sergio O. Martinez-Chapa
v
Acknowledgements
Authors would like to acknowledge the financial support of Tecnológico de
Monterrey, Mexico, for the special grant (grant number: 002EICII01) awarded by
the Sensors and Devices Focus Group, School of Engineering and Sciences,
Tecnológico de Monterrey, Monterrey, Mexico.
vii
Contents
1 Fundamentals and History of ELISA: The Evolution of the
Immunoassays Until Invention of ELISA . . . . . . . . . . . . . . . . . . . . . 1
1.1 Evolution of the Immunoassays Until Invention of ELISA . . . . . . 1
1.1.1 Side Chain Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
1.1.2 Antigen-Antibody Binding Theory . . . . . . . . . . . . . . . . . . 2
1.1.3 Discovery of Antibody Structure . . . . . . . . . . . . . . . . . . . 3
1.1.4 Invention of Radioimmunoassay (RIA) . . . . . . . . . . . . . . . 3
1.1.5 Invention of Enzyme Linked Immunosorbent Assay
(ELISA) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
1.2 Principles of the Immune System . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2.1 Antibody Production in Human Body. . . . . . . . . . . . . . . . 5
1.2.2 Different Types of Antibodies . . . . . . . . . . . . . . . . . . . . . 5
1.2.3 Antigen-Antibody Coupling . . . . . . . . . . . . . . . . . . . . . . . 10
1.3 Biomolecular Interactions Between Antibody and Antigen . . . . . . 11
1.3.1 Hydrogen Bonding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
1.3.2 Hydrophobic Interaction. . . . . . . . . . . . . . . . . . . . . . . . . . 12
1.3.3 Ionic Attraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
1.3.4 Van der Waals Forces . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2 General Overviews on Applications of ELISA . . . . . . . . . . . . . . . . . 19
2.1 Applications of ELISA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
2.1.1 Food Industry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
2.1.2 Vaccine Development . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.1.3 Immunology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.1.4 Diagnosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
2.1.5 Toxicology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
2.1.6 Drug Monitoring and Pharmaceutical Industry. . . . . . . . . . 25
2.1.7 Transplantation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
ix
x Contents
3 Step by Step with ELISA: Mechanism of Operation, Crucial
Elements, Different Protocols, and Insights on Immobilization
and Detection of Various Biomolecular Entities . . . . . . . . . . . . . . . . 31
3.1 Mechanism of Operation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
3.2 Different Elements of the Assay . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.2.1 Solid Phase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.2.2 Adsorbents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.2.3 Washing Agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
3.2.4 Blocking Agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
3.2.5 Enzymes and Substrates. . . . . . . . . . . . . . . . . . . . . . . . . . 34
3.2.6 Stopping Process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
3.2.7 Reading Techniques. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
3.2.8 Reading Apparatus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
3.2.9 Controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
3.3 Different Protocols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
3.3.1 Direct ELISA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
3.3.2 Indirect ELISA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
3.3.3 Sandwich ELISA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
3.3.4 Double Sandwich ELISA. . . . . . . . . . . . . . . . . . . . . . . . . 47
3.3.5 Competitive ELISA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
3.4 Initial Interaction of the Biomolecules with the Surface . . . . . . . . 47
3.5 Immobilization Techniques for Protein Attachment. . . . . . . . . . . . 49
3.5.1 Physical Immobilization. . . . . . . . . . . . . . . . . . . . . . . . . . 49
3.5.2 Immobilization via Entrapment. . . . . . . . . . . . . . . . . . . . . 50
3.5.3 Covalent Immobilization . . . . . . . . . . . . . . . . . . . . . . . . . 50
3.5.4 Oriented Immobilization. . . . . . . . . . . . . . . . . . . . . . . . . . 52
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
4 Evaluation of the Detection Results Obtained from ELISA . . . . . . . 57
4.1 Conducting a Reliable Assay. . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
4.1.1 Sources of Errors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
4.1.2 Troubleshooting. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
4.2 Key Parameters in ELISA Evaluation . . . . . . . . . . . . . . . . . . . . . 62
4.2.1 Sensitivity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
4.2.2 Specificity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
4.2.3 Accuracy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
4.2.4 Limit of Detection (LOD) . . . . . . . . . . . . . . . . . . . . . . . . 64
4.3 Measurable Units in ELISA . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
5 Advantages, Disadvantages and Modifications of Conventional
ELISA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
5.1 Significance of Conventional ELISA . . . . . . . . . . . . . . . . . . . . . . 68
5.2 Shortages of Conventional ELISA. . . . . . . . . . . . . . . . . . . . . . . . 68
Contents xi
5.3 Materials of Choice for Fabrication of ELISA Well Plates . . . . . . 69
5.4 Different Types of ELISA Well Plates. . . . . . . . . . . . . . . . . . . . . 70
5.5 Modified ELISA Platforms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
5.5.1 ELISA on Coated Platforms. . . . . . . . . . . . . . . . . . . . . . . 72
5.5.2 ELISpot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
5.5.3 Plasmonic ELISA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
5.5.4 Sphere-/Bead-Based ELISA . . . . . . . . . . . . . . . . . . . . . . . 80
5.5.5 Paper-Based ELISA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
5.5.6 Fiber-Based ELISA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90
5.5.7 ELISA in Micro-Devices . . . . . . . . . . . . . . . . . . . . . . . . . 98
5.5.8 Other Strategies. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108