The ELISA Guidebook: Second Edition / Edition 2

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The Enzyme Linked Immunosorbent Assay (ELISA) continues, as predicted, to be the predominant assay used in biomedical and agricultural research. While little has changed in the principles of the ELISAs used since the publication of The ELISA Guidebook, many technological advantages, including the increasing number of reagents available for exploitation and the larger spectrum of instruments to measure the coloured products of ELISAs, have greatly benefited the field. Other testing technologies using molecular techniques, predominantly the conventional and more and more often, real time PCR, are rapidly evolving to help diagnose and differentiate disease agents and identify substances present in exquisitely small amounts. The ELISA is still needed either alone in mass screening, such as serological assays, or used in tandem with molecular techniques to fully evaluate the molecular and antigenic nature of analytes.

John R. Crowther provides today’s premier practical guide to the understanding and application of ELISA. This Edition enlarges on 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. The author describes each method in great detail to ensure experimental success and includes advice on equipment choice, maintenance, and calibration. Wherever possible, helpful written explanations are provided along with copious diagrams.

With its numerous worked examples, detailed instructions, and extensive illustrations, The ELISA Guidebook, Second Edition offers a powerful synthesis of all the basic concepts and practical experimental details investigators need to understand, develop, and apply ELISA methodology successfully in day-to-day basic and clinical research.

The book contains black-and-white illustrations.

This newly revised edition continues to present the fundamental aspects of the heterogeneous enzyme linked immunosorbent assay (ELISA). Much new information has been added to further present the ELISA method to bioscientists using it for the detection and quantification of antigens and antibodies (e.g., diagnosis). Designed for the newcomer as well as the experienced practitioner this book gives numerous examples of the use of ELISA and the previous edition has been successfully used as an essential teaching guide. New chapters address monoclonal antibodies, the validation of assays, chessboard titrations and more—all of which will contribute to an improved efficiency of ELISA development.

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Editorial Reviews

Doody's Review Service
Reviewer: Ruben Mestril, PhD (Loyola University Medical Center)
Description: This is a compilation of 14 chapters on different aspects of the enzyme-linked immunosorbent assay (ELISA) with a final chapter that provides a series of test questions. The book contains detailed instructions and extensive illustrations with many practical examples on the technique permitting even investigators not familiar with the technique to learn all the basic concepts and practical experimental details needed to understand and apply ELISA methodology successfully in basic and clinical research.
Purpose: The ELISA is still one of the predominant assays used in biomedical research. Although not much has changed concerning the principles used in an ELISA, there have been many technological advances since the first edition of this guidebook. There are a number of new and better instruments to measure the final products of an ELISA. In addition, there are an increasing number of different reagents available to visualize the results of the assay. Although there are other testing technologies using molecular techniques, such as RT-PCR, that are able to detect and identify agents present in only minute amounts, the ELISA is still required for large screenings, or even in conjunction with other molecular techniques.
Audience: The book is written mainly for molecular and cellular biologists, biochemists, pharmaceutical scientists, immunologists, pathologists, laboratory medicine researchers, and all researchers using biochemical assays. The guidebook provides practical instructions that will be useful to all investigators engaged in either basic or clinical research.
Features: The guidebook provides clear instructions on how to develop any type of ELISA according to the investigator's needs. Among the most interesting features is chapter 12 on the more advanced statistical methods for quality assurance, test validation, and interpretation. This chapter examines the different statistical factors involved in assessing the validity of the results obtained by using an ELISA approach. Also of interest is chapter 15 that provides a series of test questions covering the material presented in the first 14 chapters. This permits the use of the guidebook as a textbook for a course on ELISA methodology.
Assessment: This volume of Methods in Molecular Biology is an excellent manual for the understanding and development of the ELISA. The guidebook covers all the relevant points necessary for investigators to develop a useful ELISA for their particular needs. This second edition should prove to be a valuable companion for all researchers in the field of basic and clinical research.
Crowther (International Atomic Energy Agency, Vienna) expands the information about the enzyme-linked immunosorbent assay (ELISA) from his 1995 . He reorganizes the chapters and incorporates new information, particularly dealing with the chessboard titration of reagents, quality control, monoclonal antibodies, validating assays, statistics, and epidemiological considerations. Though he writes primarily for scientists with previous experience of the technique, his guide could also be helpful for newcomers to it and as a teaching tool<-->he includes test questions for each chapter. The plastic comb binding lies flat. Annotation c. Book News, Inc., Portland, OR (
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Product Details

  • ISBN-13: 9781603272537
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 12/12/2008
  • Series: Methods in Molecular Biology Series , #516
  • Edition description: 2nd ed. 2009
  • Edition number: 2
  • Pages: 566
  • Product dimensions: 7.70 (w) x 10.30 (h) x 1.20 (d)

Table of Contents

1) Overview of ELISA in Relation to Other Disciplines

2) Systems in ELISA

3) Stages in ELISA

4) Titration of Reagents

5) Theoretical Considerations

6) Practical Exercises

7) Monoclonal Antibodies

8) Validation of Diagnostic Tests for Infectious Diseases

9) Charting Methods for Internal Quality Control for Competition ELISA

10) Charting Methods for Internal Quality Control of Indirect ELISA

11) Ruggedness and Robustness of Tests: Aspects of Kit Use and Validation

12) More Advanced Statistical Methods for Quality Assurance, Test Validation and Interpretation

13) Internal Quality Control and External Quality Management of Data in Practice

14) Immunochemical Techniques

15) Test Questions

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