Description
Experimental Immunology aims to help learners in their research exercise. This book elaborates on the methodology for isolation of antigens, production of monoclonal & polyclonal antibodies, which are the basic milestones in Immunology. Immunology is essential for the study of pathogens and remains as the cardinal
tool for diagnosis. A separate chapter explicates the applications of immunological tools. Experiments in Immunology are not complete without experimental animals and hence immunization and handling of experimental animals are given due importance in this book.
Table of Content
Foreword
Preface
Acknowledgements
Abbreviations
Chapter 1 Antigens
1.1 Antigens –An Overview
1.1.1 Molecules as Antigens
1.1.2 Synthetic Antigens
1.1.3 Complex Antigens
1.2 Isolation and Identification of Bacterial Antigens
1.2.1 Introduction
1.2.2 Types of Bacterial Antigens
1.2.3 Methods for Isolation of Bacterial Antigens
1.2.4 Isolation of Antigens
1.2.5 Free Protein Antigens
1.2.6 Bound Protein Antigens
1.2.7 Carbohydrate Antigens
1.2.8 Appendages as Antigens
1.2.9 Mycobacterial Antigens
1.2.10 Recommendations/Suggestions for Preparation of Bacterial Antigens
1.3 Preparation of Viral Antigens
1.3.1 Introduction
1.3.2 Types of Viral Antigen
1.3.3 Major Classes of Viral Antigens
1.3.4 Source for Virus
1.3.5 Uses of Viral Antigens
1.3.6 Antigens for use as Immunogens
1.3.7 Purification Procedure
1.4 Preparation of Fungal Antigens
1.4.1 Introduction
1.4.2 Fungal Antigens
1.4.3 Growth Conditions
1.4.4 Crude Antigenic Extracts
1.4.5 Exo antigens
1.4.6 Fungal Allergen Preparations
1.4.7 Polysaccharide Antigens
1.4.8 Polysaccharide free Protein Antigens Preparation
1.5 Preparation of Antigens of Parasites
1.5.1 Introduction
1.5.2 Complexity of Parasites
1.5.3 Monoclonal Antibodies for Parasite Antigens
1.6 Preparation of Synthetic Antigens
1.6.1 Introduction
1.6.2 Polypeptidyl Proteins
1.6.3 Linear Polymers
1.6.4 Multi chain Polymers
Multiple choice questions
Chapter 2 Immunoglobulins
2.1 Production of Polyclonal Antibodies
2.1.1 Strategy for Polyclonal Antiserum Production
2.2 Production of Murine Monoclonal Antibodies
2.2.1 Basic Principles for B-Hybridoma Production
2.2.2 Strategy for Murine Monoclonal Antibody Production
2.2.3 Applications of Monoclonal Antibodies
2.2.4 Human Monoclonal Antibodies
2.3 Immunochemical Analysis of Immunoglobulins
2.3.1 Introduction
2.3.2 Structural Relationships
2.3.3 Properties of Immunoglobulins (Igs)
2.3.4 Major Immunoglobulin Classes
2.3.5 Immunoglobulin Subclasses
2.3.6 Chemical Characterization of Igs
2.3.7 Analysis of Immunoglobulin Subunits
2.4 Characterization of Monoclonal Antibodies
2.4.1 Introduction
2.4.2 Purification of Monoclonal Antibodies
2.4.3 Hybridoma Supernatant
2.4.4 Serum free Medium
2.4.5 Characterization of Monoclonal Antibody
2.4.6 Binding Assays
2.5 Determination of Three-dimensional Structures of Immunoglobulins
2.5.1 Introduction
2.5.2 Crystallization of Igs
2.5.3 Three-dimensional Structures of Igs
Multiple choice questions
Chapter 3 Antigen Antibody Interactions
3.1 Principle of Antigen Antibody Interactions
3.2 Factors that Affect Antigen Antibody Reactions
3.3 In vitro Antigen Antibody Reactions
3.3.1 Precipitation Reaction
3.3.2 Agglutination Reaction
3.3.3 Complement Fixation Reaction
3.3.4 Immunoassays
3.4 Immunoprecipitation
3.5 Immunofluorescence
3.5.1 Applications of Immunofluorescence Technique
3.6 Immunohistochemistry
3.6.1 Preparation of Tissues or Cells
3.6.2 Preparation of Solid Tissue for Paraffin Embedding
3.6.3 Preparation of Fluorochrome-Conjugated Antisera
3.7 Flow-Cytometry
3.8 Immunoelectron Microscopy
Multiple choice questions
Chapter 4 Applications of Immunodiagnostic Methods
4.1 Principles of Immunodiagnostic Methods
4.1.1 Features of Humoral Immune Response Useful in Diagnostic Testing 4.1.2 Principles of Serological Test Methods
4.2 Application of Immunological Techniques in Bacteriology
4.2.1 Learning Objective
4.2.2 Agglutination
4.2.3 Immobilization Tests
4.2.4 Complement Fixation Tests
4.2.5 Immune Hemolysis
4.2.6 Precipitation
4.2.7 Toxin Neutralization Test
4.2.8 Fluorescent Antibody Technique
4.2.9 Immuno electron Microscopy
4.2.10 Enzyme Linked Immunosorbent Assay (ELISA)
4.3 Applications of Immunologicals Methods in Virology
4.3.1 Principle
4.3.2 Humoral Immunity
4.3.3 Virus Neutralization
4.3.4 Precipitation
4.3.5 Immunofluorescence
4.3.6 Immune Electron Microscopy
4.3.7 ELISA and Radiommunoassay
4.3.8 Cellular Immunity
4.3.9 Direct Cell Cytotoxicity
4.3.10 Uses of Immunological Methods of Virology
4.4 Applications of Immunological Methods in Mycology
4.4.1 Introduction
4.4.2 Immunological Methods
4.4.3 Applications of Immunological Methods
4.5 Applications of Immunological Methods in Helminthology
4.5.1 Applications
4.5.2 Production of Helminthes Antigens
4.5.3 Serological Diagnosis
4.6 Application of Immunological Methods in Protozoology
4.6.1 Immunodiagnosis – The Recognition of Infected Hosts
4.6.2 Immunocharacterization – The Recognition of Subspecific Population of
Parasitic Protozoa
4.6.3 Immunization
4.6.4 Protozoological Consideration of Life Cycles
4.6.5 Production of Important Protozoal Genera for Immunological Study Multiple choice questions
Chapter 5 Experimental Animals
5.1 Immunization of Experimental Animals
5.1.1 Introduction
5.1.2 Antigen Dose
5.1.3 Control of Immune Response
5.1.4 Immunopotentiation
5.1.5 Basic Procedures for Immunization
5.1.6 Preparation and use of Common Adjuvants
5.1.7 Immunization in Other Experimental Animals
5.2 Handling of Experimental Animals
5.2.1 Handling Methods
5.2.2 Administration of Inoculum
5.2.3 Bleeding of Experimental Animals
Multiple choice questions
Appendix
Glossary
Bibliography
Index
About The Author
Dr. V. Mahalakshmi is currently heading the department of Microbiology at Madras Christian College, Chennai. With over two decades of teaching experience, her doctoral work focused on Polyethylene biodegradation. The author firmly believes that the World of Microbiology is fascinating and challenging and she endeavours to laboriously discover its intriguing world. She is part of many professional bodies associated with the branch of Microbiology. This book is second in her academic career.
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