T. T. Ngo and H. M. Lenhoff Department of Developmental and Cell Biology University of California, Irvine, CA 92717 In 1959, Yalow and Berson used insulin labeled with radioactive iodine to develop a quantitative immunological method for determining the amount of insulin in human plasma. Their method depends upon ~ competition between insulin labeled with radioactive iodine (II 1) and unlabeled insulin from plasma for a fixed and limited number of specific binding sites on the antibody to insulin. The amount of the labeled insulin bound to the antibody is inversely proportional to the amount of insulin in the plasma sample. Their method, which is so elegantly simple in concept, is made possible by the ability to detect with ease extremely low levels of radioactivity, and by the exquisite specificity of an antibody capable of specifically binding the analyte. Such a combination of sensitivity and specificity is the basis of this versatile analytical tool called radioimmunoassay (RIA). Twelve years later, Engvall and Perlmann (1971) and Van Weemen and Schuurs (1971) independently introduced the use of enzymes as another category of sensitive and even more versatile labels for use in immunoassays. Engvall and Perlmann (l971) coined the term ELISA, which stands for Enzyme Linked Immunosorbent Assay.

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.

Immunoassays are among the most powerful and sensitive technologies now available for patient diagnosis and monitoring. This book is an indispensable guide to information on the theory and practice of immunoassays. It discusses the scientific basis of these technologies in a logical, organized, and heuristic manner and provides protocols for specific assays. The contents of this unique book are balanced among theory, practical issues, quality control, automation, and subspecialty areas, making it ideal for health science students, laboratory scientists, and clinicians. Presents up-to-date information Provides extensive cross-referencing Covers theory and practice in full detail Written by leading authorities

This unique reference provides a pragmatic approach to the development of successful commercial immunodiagnostic products based on enzyme immunoessay technology. Presenting both the basic and applied principles, Enzyme Immunoassays gathers information on all aspects of this process, from the initial conceptualization to the introduction of the product to the market.

The basis of all immunoassays is the interaction of antibodies with antigens. The most widely used immunoassay technique is radioimmunoassay (RIA) which was first developed by Yalow and Berson in 1959. The principle of RIA is elegantly simple. It utilizes a competitve binding reaction between analytes and a radio-labeled analog of the analytes (the tracer) for anti-analyte antibodies. In addition to its exquisite specificity, extraordinary sensitivity, good accuracy and precision, ease and rapidity of assay and simplicity of assay development, the applicability of RIA to a wide variety of substances has made it one of the most powerful and versatile analytical methods of the 20th century and beyond. Millions of RIA's are being performed annually on clinical, biological and environmental samples in licensed laboratories. In order to expand the use of RIA beyond the confines of these laboratories to areas like physician's offices, patients' homes, economically less developed countries, agricultural fields, large scale and continuing screening tests for infectious diseases, it has become necessary to develop non-isotopic labels. Indeed the last fifteen years have seen the development of a great number of ingenious non-isotopic labels in immunoassay so that a whole new industry capitalizing on the potential market for non isotopic immunoassays has appeared. It is the purpose of this volume to present in depth, state-of-the-art reviews on techniques used in non-isotopic immunoassays. Topics covered include: (1) Enzyme-labeled immunoassay; (2) Luminescene immunoassay; (3) Immunoassay at liquid-solid interface; (4) Membrane immunoassay and (5) "Particle"-mediated immunoassay.

The purpose of this book is to focus attention on some of these ideas and concepts. In doing so, it has captured a glimpse of the past and it attempts a projection of the future, but mostly it reveals an overview of the field as it exists as the present time. It aims to serve to spawn further growth in ideas and encourage applications to increasingly broader segments of both clinical and general analytical chemistry fields.