
Immunodiagnostics
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Content
- Intro
- Contents
- List of contributors
- Abbreviations
- 1. Principles of immunodiagnostic tests and their development
- with specific use of radioisotopes as tracers
- 1. Introduction
- 2. Classification of type
- 3. Selection and preparation of reagents
- Antibody or antiserum
- Selection of antibody or antiserum
- Purification of antibodies and antisera
- Selection of label
- Preparation of [sup(125)]I-radioiodinated tracers
- Purification of [sup(125)]I-radioiodinated tracers
- Selection and preparation of standard or calibrant
- 4. Assay design
- Assay format
- General reagents and additives
- Sample preparation
- Optimal protocol and concentration of reagents
- Assay validation
- Appendix
- References
- 2. Solid-phase supports
- 1. Introduction
- 2. Solid phase supports
- Polystyrene
- Other organic polymers
- Cellulose particles
- Agarose, sephacryl and sephadex particles
- Magnetizable particles
- Membranes
- 3. Preparation of solid supports
- 4. Passive adsorption
- 5. Covalent coupling
- Derivatization of solid supports
- Covalent coupling reactions with solid supports
- 6. Indirect coupling methods
- Indirect coupling of antibody via a second antibody
- Indirect coupling of antibody via binding proteins (A and G)
- Indirect binding of antibody via the biotin-avidin interaction
- Indirect binding of antibodies labelled with a hapten (fluorescein)
- 7. Blocking
- 8. Storage
- 9. Assay wash solutions
- Appendix A: Suppliers of solid supports
- Appendix B: List of suppliers
- References
- 3. Enzyme-labelled tests with colorimetric, fluorimetric and chemiluminescent detection systems
- 1. Introduction
- 2. Selection of enzyme-labels
- 3. Enzyme preparation and assignment of enzymatic activity
- Preparations
- Enzymatic assays
- 4. Conjugation methods
- Activation of small molecules and their covalent coupling to enzymes
- Activation of glycoprotein enzymes and their coupling to proteins
- Activation of glycoproteins and their covalent coupling to enzymes
- Coupling via bifunctional linking molecules
- 5. Purification, storage and testing of enzyme conjugates
- Gel filtration of conjugate
- Lectin chromatography of glycoprotein enzyme conjugates
- Fractionation of antibody conjugates by Protein A chromatography
- Stabilization and storage of enzyme conjugates
- Testing of conjugates
- 6. Signal generation
- Spectrophotometric systems for solid phase enzyme-labelled assays
- Enzyme amplification
- Immunostaining procedures
- Fluorescent systems
- Enhanced chemiluminescent systems
- References
- 4. Time-resolved fluorescence immunoassay
- 1. Introduction
- 2. Principles of time-resolved fluorescence
- 3. Protein labelling with lanthanide chelates
- 4. Storage of labelled proteins
- 5. Application to two-site immunometric assay of proteins
- 6. Application to competitive binding immunoassays of haptens
- 7. Europium-labelled hapten assay
- 8. Monoclonal antibody screening procedures
- 9. Novel research assays
- 10. Non-separation time-resolved fluoroimmunoassay
- 11. Simultaneous immunoassays
- 12. Conclusion
- Acknowledgements
- Appendix: List of suppliers
- References
- 5. Light scattering techniques
- 1. Introduction
- 2. Fundamental aspects of immunoaggregation
- Monitoring techniques
- End-point versus rate measurement
- Analyte and assay format
- Antigen excess
- Optimization strategies
- Validation strategies
- 3. Non-enhanced methodology
- Antibody selection
- Choice of buffer and PEG concentration
- Assay performance
- 4. Enhanced methodology
- Choice of assay format
- Selection of particle
- Coupling chemistry
- Evaluation of particle reagents
- Antibody selection and optimization
- Buffer optimization
- Assay performance
- 5. Summary
- References
- Acknowledgements
- 6. Enzyme amplification: A means to develop fast ultrasensitive immmoassays
- 1. Introduction
- 2. Practical applications
- Water and buffers
- Colorimetric enzyme amplification
- 3. Fluorimetric enzyme amplification
- 4. Conclusions
- Appendix: List of suppliers
- References
- 7. Equipment and automation
- Appendix on dose-response curve fitting
- 1. Introduction
- Features of automation
- 2. Steps in immunoassay analysis
- Sample identification-bar-coding
- Worklist generation
- Transfer of samples to reaction vessel
- Addition of immunoreagents
- Incubation
- Separation where necessary
- Signal generation
- Signal detection
- Curve fitting-response calibration
- Quality control
- Report generation
- Export of results to LIMS
- 3. Equipment
- Bar code readers
- Worklist generators
- Pippettors/diluters
- Reagent dispensers
- Incubators/shakers
- Washers
- Signal detectors
- Response curve calibration software
- Quality control
- Sample/reagent processors
- Assay processors
- Robotic arms
- Fully automated analysers
- Clinical chemistry analysers
- Cleaning and disinfection
- Safety features
- 4. Analytical considerations
- 5. Conclusions
- References
- Appendix
- 8. Quality assurance in immunodiagnostics
- 1. Introduction
- The need for quality
- The purpose of quality assurance
- Implementation of a quality assurance programme
- 2. Quality systems
- Quality assurance (QA)
- Internal quality control (IQC)
- External quality assessment (EQA)
- Audit
- Accreditation of laboratories
- Validation of results
- Good manufacturing practice (GMP)
- Training and education
- Evaluation of reagents and equipment
- 3. Quality assurance
- Test selection and siting
- Pre-analytical quality assurance
- Post-analytical quality assurance
- Non-analytical quality assurance
- Summary
- 4. Internal quality control
- Simple IQC procedures
- Statistical quality control techniques
- Validated control techniques
- Techniques using patients' data
- Assay validation
- Clinical validation
- Corrective action
- Summary
- 5. External quality assessment
- The purpose of EQA
- Practical considerations
- Scheme design
- Competition in EQA
- How to use EQA participation
- Selection of an EQA scheme
- 6. Materials for IQC and EQA
- Material sources and preparation
- Material presentation
- Summary
- 7. Discussion
- Why use QA at all?
- What is good enough?
- What reference intervals should be used?
- Who has responsibility for quality?
- Future prospects
- Acknowledgements
- References
- List of suppliers
- Index
- A
- B
- C
- D
- E
- F
- G
- H
- I
- K
- L
- M
- N
- O
- P
- Q
- R
- S
- T
- U
- V
- W
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