The Elisa Guidebook: Second Edition

ISBN-13: 9781617378843 / Angielski / Miękka / 2010 / 566 str.

John R. Crowther

Opis

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.

Recenzje

From the reviews of the second edition:

"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 ... and apply ELISA methodology successfully in basic and clinical research. ... The book is written mainly for molecular and cellular biologists, biochemists, pharmaceutical scientists, immunologists, pathologists, laboratory medicine researchers, and all researchers using biochemical assays. ... This volume ... is an excellent manual for the understanding and development of the ELISA." (Ruben Mestril, Doody's Review Service, May, 2009)

"It is the practical ELISA bible, a definitive manual covering theoretical aspects, commercial reagents, troubleshooting, instrumentation, enzyme-linked conjugates and microwell plates. ... Throughout this book excellent use is made of flow diagrams, graphs and tables to aid the reader. ... The ELISA Guidebook is a must for institutional libraries, departments of immunology, hospital diagnostic laboratories, research laboratories using ELISA-based assays and companies developing ELISA immunodiagnostic kits. The book is well worth its price." (Cyril J. Smyth, Microbiology Today, November, 2009)

"The 2nd Edition of The ELISA Guidebook is essentially the author's tribute to the ELISA technology that has revolutionised all laboratories ... . This edition well lives up to its name ... . Overall, I rate this book as an extremely valuable resource ... to the daily ELISA user. I recommend this book to all Biomedical Scientists, Clinical Scientists and trainees in Laboratory Medicine and that one copy should be available in every laboratory that makes extensive use of ELISA." (Sujoy Khan, Immunology News, November, 2009) "The book describes the methods involved in heterogeneous enzyme-linked immunosorbent assays (ELISA) that provide ideal systems for dealing with a wide range of studies in many biological areas. ... This method is widely using in various scientific and industrial areas ... . provides a premier practical guide to the understanding and application of ELISA. Each method is described in great detail, and the whole book is an excellent manual for many researchers in biochemistry, molecular biology, immunochemistry, biotechnology, and medicine." (G. Ya. Wiederschain, Biochemistry, Vol. 9, 2009)

Spis treści

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

Blurb

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.