The ELISA Guidebook

1. Front Matter

   Pages i-xv

   PDF

2. Overview of ELISA in Relation to Other Disciplines

   Pages 1-8

3. Systems in ELISA

   Pages 9-42

4. Stages in ELISA

   Pages 43-78

5. Titration of Reagents

   Pages 79-109

6. Theoretical Considerations

   Pages 111-147

7. Practical Exercises

   Pages 149-224

8. Monoclonal Antibodies

   Pages 225-289

9. Validation of Diagnostic Tests for Infectious Diseases

   Pages 291-334

10. Charting Methods for Internal Quality Control for Competition ELISA

    Pages 335-380

11. Charting Methods for Internal Quality Control of Indirect ELISA

    Pages 381-429

12. Ruggedness and Robustness of Tests: Aspects of Kit Use and Validation

    Pages 431-465

13. More Advanced Statistical Methods for Quality Assurance, Test Validation, and Interpretation

    Pages 467-516

14. Internal Quality Control and External Quality Management of Data in Practice

    Pages 517-541

15. Immunochemical Techniques

    Pages 543-554

16. Test Questions

    Pages 555-562

17. Back Matter

    Pages 563-566

    PDF

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.

• Antigen
• Diagnostic techniques
• ELISA
• Enzyme linked immunosorbent assays
• PCR
• Quality control
• infectious disease
• antibodies

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) thatprovide 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)

• International Atomic Energy Agency, Animal Production & Health Section, Vienna, Austria

  John R. Crowther

Policies and ethics