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.

"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.

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.

John R. Crowther provides today's premier practical guide to the understanding and application of ELISA. Updating and greatly expanding his widely appreciated earlier publication, ELISA Theory and Practice (1995), this important work introduces chapters on such major new topics as checkerboard titrations, quality control of testing, kit production and control, novel monoclonal antibodies, validation of assays, statistical requirements for data examination, and epidemiological considerations. With its numerous worked examples, detailed instructions, and extensive illustrations, The ELISA Guidebook offers a powerful synthesis of all the basic concepts and practical experimental details investigators need to understand, develop, and apply the new ELISA methodology successfully in day-to-day basic and clinical research.

This book will serve as a primer for both laboratory and field scientists who are shaping the emerging field of molecular epidemiology. Molecular epidemiology utilizes the same paradigm as traditional epidemiology but uses biological markers to identify exposure, disease or susceptibility. Schulte and Perera present the epidemiologic methods pertinent to biological markers. The book is also designed to enumerate the considerations necessary for valid field research and provide a resource on the salient and subtle features of biological indicators.

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.

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.