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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.
There have been very few developments that markedly affect the need to greatly revise the text from the last version of this book. This is testament to the fact that hetero- neous enzyme-linked immunosorbent assays (ELISA) provide ideal systems for dealing with a wide range of studies in many biological areas. The main reason for this success is test flexibility, whereby reactants can be used in different combinations, either attached passively to a solid phase support or in the liquid phase. The exploitation of the ELISA has been increased through continued development of specifically produced reagents, for example, monoclonal and polyclonal antibodies and peptide antigens coupled with the improvement and expansion of commercial products such as enzyme-linked conjugates, substrates and chromogens, plastics technology and design of microwell plates, inst- mentation advances and robotics. However, the principles of the ELISA remain the same. There has been some rearrangement of chapters plus addition of three new ones dealing with charting methods for assessing the indirect ELISA, ruggedness and robustness of tests-aspects of kit use and validation, and internal quality control and external quality management of data, respectively. These reflect the need to control what you are doing with ELISA and to exploit the method to its full extent. I do not apologize for dealing with the same areas in different ways a number of times, as it is imperative that principles are understood to allow planning, operation, and control of ELISA.
ELISA: Theory and Practice introduces to scientists at all levels of expertise the principles of the most commonly used assay technique known as the Enzyme Linked Immunosorbent Assay. The book provides readers with full descriptions of the basic systems that make ELISA one of the most powerful techniques in science today, and also examines in detail the data obtained by ELISA and their analysis and actual manipulation. ELISA: Theory and Practice is designed not only to train novices in the science of ELISA, but also to aid investigators experienced in any of the biological sciences in performing independently assays of antibodies and antigens. Mastery of the book's contents will allow readers to fully appreciate exactly how and why assays function, as well as permit the efficient development of individual assays that are both rapid and accurate.
The last few years have seen the rapid development of new methodology in the field of molecular biology. New techniques have been regularly introduced and the sensitivity of older techniques greatly improved upon. Developments in the field of genetic engineering in particular have contributed a wide range of new techniques. The purpose of this book therefore is to introduce the reader to a selection of the more advanced analytical and preparative techniques which the editors consider to be frequently used by research workers in the field of molecular biology. In choosing techniques for this book we have obviously had to be selective, and for the sake of brevity a knowledge of certain basic biochemical techniques and terminology has been assumed. However, since many areas of molecular biology are developing at a formidable rate and constantly generating new terminology, a glossary of terms has been included. The techniques chosen for this book are essentially based on those used in a series of workshops on 'techniques in molecular biology' that have been held at The Hatfield Polytechnic in recent years. In choosing these chapters we have taken into account many useful suggestions and observations made by participants at these workshops. Each chapter aims to describe both the theory and relevant practical details for a given technique, and to identify both the potential and limitations of the technique. Each chapter is written by authors who regularly use the technique in their own laboratories.
This volume is a practical biochemical guide to the Enzyme-Linked Immunosorbent Assay (ELISA), used to detect a target substance in a liquid sample. The ELISA is an important and widely used diagnostic tool in medicine, animal health, botany and quality assurance processes in food and beverage production. An introductory chapter orients the reader on the basic structure and function of immunoglobulins and their fragments while subsequent chapters outline the methodology to generate monoclonal antibodies using hybridoma technology and the general methods used to purify antibodies. Multiple chapters demonstrate how to creatively use the properties of the antibody to identify, localize and quantify target analytes to answer questions and resolve problems. The reader will learn how to use a variety of immunoassay strategies, reporters and detection systems that will undoubtedly facilitate their efforts to gain answers to their own questions. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, ELISA: Methods and Protocols seeks to provide both professionals and novices with the technical information necessary for the reader to successfully use the immunoassay as part of the discovery process.
This book shows the various sandwich assays that are constructed from recognition molecules, such as antibodies, oligonucleotide sequences and aptamers, developed as a result of nano- and biotechnology advances. It consists of ten chapters presenting interesting examples of these assays, organized according to the type of analytic methods (colorimetric, fluorescence, electrochemical, etc.) and detected objects (protein, nucleic acid, small-molecule, ion, etc.). It also includes a chapter discussing the introduction of sandwich assays as biosensors for the detection of a range of targets. It is an interesting and useful resource for a wide readership in various fields of chemical science and nanotechnology.
The book appeared in two previous Slovak editions for university students in Czechoslovakia. This edition presents a completely new version updated according to recent advances not only in immunochemistry and essential immunology but also in molecular biology, biochemistry and molecular genetics. The scope of the book is considerable since the goal was to cover the field of immunochemistry from the widest point of view including both the topic and methods of contemporary immunochemistry. Each chapter provides basic information on a specific subtopic, clearly and understandab ly, and presents principles of individual immunochemical methods. I am confident that the book will fill the gap between the books on essential immunology and highly specialised books on individual areas of immunoche mistry (e. g. on antibodies, antigens, numerous immunochemical techniques, etc. ). It may also prove useful for beginning investigators from different biological and medical fields as it supplies basic information needed for solving their scientific problems by immunochemical approaches. I do hope that readers will find the text stimulatory and pleasury to read. I wish to thank all colleagues and friends for supplying their own results, suggestions and for their encouraging comments. My thanks go also to the editors and publishers for their valuable contribution to the preparation of the book. 1 Introduction The term immunochemistry was coined by the Swedish chemist ARRHENIUS who used it for the first time in his lectures in 1907.
Structure of Antigens discusses a variety of topics dealing with the structural basis of antigenicity. Topics include the analytical methods used to elucidate the structure of antigens, the structure of antibodies, the principles underlying modern immunoassays and the measurement of antibody binding affinity, and physicochemical principles and methodological aspects. The book also considers major groups of antigens distinguished by their functional activity and biological role (e.g., drugs, autoantigens, snake toxins, allergens) or by their association with particular biological systems (e.g., antigens of microorganisms). Structure of Antigens will provide a current, useful, reference for researchers and graduate students in all fields of biological science who need an overview of antigenic specificity. VOLUME 1
This work provides rapid access to focused information on topics of Immunotoxicology not only for scientists and those dealing with laboratory aspects but also for lecturers and advanced students. Over 200 contributing authors – including many of the world’s top specialists – have contributed full essays on all relevant topics, supplemented by keyword definitions of related terms. Full essays are structured uniformly to provide reader-friendly information on all aspects of Immunotoxicology, including methods of testing and analysis, characteristics of substances, the regulatory environment and the relevance of these to humans. The single A–Z format of both types of entry makes this reference book very easy to use. The Encyclopedic Reference of Immunotoxicology is intended to be a comprehensive work of reference which will provide easy access to relevant information in the fast-growing field of Immunotoxicology.
Immunochemistry of Solid-Phase Immunoassay fills a niché in the field of immunoassay and immunology. Although solid-phase immunoassay constitutes a major technology in biology and medicine, there is no comprehensive source devoted to the immunochemical principles involved. As a result, this book will benefit students, technicians, and researchers who use this technology, as well as immunodiagnostic and biotech companies who develop the technology. The book is not a methods manual; instead, it incorporates the concepts, data, and opinions of more than 25 investigators working in this field. Topics discussed include: the chemistry of solid-phases, the behavior or antibodies and antigens on solid phases, membrane solid-phases, reaction kinetics, antigen quantitation, enzyme systems, photophysics, immunochemical considerations in data analyses, multianalyte assays and occupancy concepts, antibody quantitation, streptavidin, a review of data analysis software, and solid-phase peptide immunoassay.