Enzyme-Linked Immunosorbent Assay (Elisa): From A to Z (Paperback, 2018)

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