Assay Development and Technologies (2nd Edition) : Market Research Report

Assay Development and Technologies (2nd Edition)

Published by: Drug and Market Development Publishing

Published: Sep. 1, 2006 - 282 Pages

Chapter 1: Executive Summary: Purpose and Scope; Summary; Additional Information
Chapter 2: Introduction: Definitions; Brief Historical Perspective; Basic Issues; Drug Development and Manufacturing; Miniaturization
Chapter 3: Common Assays: Quantitation of Therapeutics and Other Compounds; Microbial Contamination; Binding; Enzyme Assays; G protein-coupled receptors; Ion Channels; Toxicology and Pharmacology; Genetic Polymorphism
Chapter 4: General Assay Design: Universal Considerations; Nature of Analyte and Matrix; Assay Objectives
Chapter 5: Format: Homogeneous and Heterogeneous; Direct and Indirect; Agonists and Antagonists; In vitro and in vivo
Chapter 6: Readout (Reporting Format): Colorimetric and Fluorometric; Calibrations and Data Analysis; Radiometric; Biological Growth; Other Readouts
Chapter 7: Validation: Installation and Operation Qualification; Performance Qualification; Scale-up
Chapter 8: In Vitro Assays: General; Enzyme Assays; Binding; Antibodies and Immunoassays; Nucleic Acids
Chapter 9: Cell-based Assay Application Areas and Associated Assays Available: General; Fixed Cells; Whole Cells; Flow Cytometry; Lysates; Microscopy
Chapter 10: Automation Platforms: Miniaturization; Robotics; Liquid Handlers; Cell Handling; Microplate Equipment
Chapter 11: Emerging Technologies: Image Analysis; Higher Density Formats; New Technologies
Chapter 12: Information Management: Data Analysis; Statistics
Chapter 13: Appendices: Appendix 1: Resources for Detailed Protocols; Appendix 2: Checklist for General Assay Development; Appendix 3: Microarray Assay Checklist; Appendix 4: Vendors; Appendix 5: Examples of Troubleshooting
Chapter 14: References
TABLE OF EXHIBITS: Exhibit 2.1. Common Causes of Non-reproducibility in Microplates; Exhibit 3.1. General Assay Formats for Binding of Labeled Ligand; Exhibit 3.2. Common FRET Donor-quencher Pairs and their Minimum Förster Radius (R0); Exhibit 3.2.. Summary of Mainstream Technologies for Ion Channel Interrogation; Exhibit 4.1. Major Assay Design Considerations and Potential Problems Addressed; Exhibit 4.2. Common Causes of Assay Non-reproducibility; Exhibit 4.3. Sample Standard Curve with Data Interpolation, Extrapolation; Exhibit 4.4. Different Assay Emphasis at Various Drug Development Stages; Exhibit 5.1. Comparison of Heterogeneous and Homogeneous Assay Formats; Exhibit 5.2 Comparison of Adsorption and Covalent Coupling in Heterogeneous Assays; Exhibit 5.3. Comparison of In Vitro and In Vivo Assays; Exhibit 5.4. List of Common In Vivo Whole Animal Assays; Exhibit 5.5. Comparison of Cell-based Assays and In Vitro Assays; Exhibit 5.6. Common Cell Lines Used for Assays; Exhibit 5.7. Common Primary Cultures for Assay; Exhibit 5.8. Common Coatings for Cell Culture Based Assays; Exhibit 6.1. Advantages and Disadvantages of Fluorescent Formats for HTS; Exhibit 6.2. General Factors Known to Perturb GFP Fluorescence; Exhibit 6.3. Conditions Tolerated by Aequorea GFP.(wt); Exhibit 6.4. Common Sources of Autofluorescence; Exhibit 6.5. Comparative Analysis of Photobleaching in Select GFP Variants; Exhibit 6.6. Examples of Light Sources for Fluorescence Excitation; Exhibit 6.7. Examples of Laser Lines Used in the Excitation of Fluorescent Proteins; Exhibit 6.8. Excitation Spectrum and Laser Lines; Exhibit 6.9. Parameters to Consider when Choosing a.CCD Camera; Exhibit 6.10. Instrument Detection Limits for Fluorescein; Exhibit 6.11 Advantages of Fluorimetry; Exhibit 6.12 Factors to Consider when Making Fluorimetric Measurements; Exhibit 6.13 Tips for Fluorescence Microplate Assays; Exhibit 6.14 Some Variables Among Fluorescence Microplate Readers; Exhibit 6.15 Sample Thickness and Microscopy Applications; Exhibit 6.16 Advantages of Two-Photon Microscopy over Confocal Microscopy; Exhibit 6.17 Requirements for Measurement of GFP Molar Extinction Coefficients; Exhibit 6.18 Molar Extinction Coefficients for Selected GFP Variants; Exhibit 6.19 Some Considerations when Measuring GFP Quantum Yields; Exhibit 6.20 Confocal Imaging Software Features; Exhibit 6.21 Typical DELFIA Assay Steps; Exhibit 7.1. Example of Factorial Design; Exhibit 7.2. Common Problems Arising from Automation of an Assay; Exhibit 8.1. Comparison of Protein Assays Based on Light Absorbance; Exhibit 8.2. Comparison of Dye Binding Protein Assays; Exhibit 8.3. Comparison of Metal Reduction Protein Assays; Exhibit 8.4. Binding Curves Plotted as Hyperbola and as Sigmoid; Exhibit 8.5. Useful Blocker Solutions; Exhibit 8.6. Comparison of Different ELISA Formats; Exhibit 8.7. Antigen Capture ELISAs Diagrammed, Direct and Indirect Formats; Exhibit 8.8. Sandwich ELISA and Competitive RIA Diagrammed; Exhibit 8.9. Recommended Number of PCR Cycles by Template Amount; Exhibit 8.10 PCR Primer Selection Criteria; Exhibit 9.1. Cell-based Assay Application Areas and Associated Assays Available; Exhibit 9.2. Common Pitfalls in Cell-based Assays; Exhibit 9.3. Minimal Controls Required for Cell-based Assays; Exhibit 9.4. Troubleshooting FISH and IFA; Exhibit 9.5. Measurable Biological Endpoints for Whole Cell Assays; Exhibit 9.6. Sample ELISpot Protocol; Exhibit 9.7. Suitable P450 Substrates; Exhibit 9.8. Classical Inducers for Rat and Human Plated Hepatocytes; Exhibit 9.9. Assayable Parameters for Vitality; Exhibit 9.10. Assayable Markers for Apoptosis; Exhibit 9.11. Common Applications for Flow Cytometry Assay; Exhibit 9.12. Dye Selection for Flow Cytometry; Exhibit 9.13. Controls Required for Flow Cytometry; Exhibit 11.1. Assays Suitable for Development Using HCS; Exhibit 11.2. Multiplexed Assay Concept; Exhibit 11.3. Comparison of Different Multiplexed Assay Technologies; Exhibit 12.1. Examples of Kinetic Constants for Fast Enzymes; Exhibit 12.2. Ordered Steps for Quality Control of UHT Data

Abstract

D&MD’s new professional development publication—Assay Development& Technologies, 2nd Edition—offers essential information to speed and simplify assay development. This professional development guide thoroughly evaluates the capabilities, strengths, weaknesses, and expectations of leading assays and assay technologies to provide you with the information necessary to select and develop the appropriate assay for the task at hand.

Purpose and Scope

The purpose of this report is to help biotechnology and pharmaceutical professionals choose and develop appropriate assays. This Second Edition includes additional content to reflect recent advancements in assay technology. A presentation of the many applications for whole cell assays as well as an expanded discussion of assay multiplexing has been added. This material recognizes the growing importance of high content, cell-based assays in drug discovery and candidate development. The section on assay methods for ion channels has also been expanded due to the emergence of analytical platforms for high-throughput screening of this very significant pharmaceutical target class. Finally, a lengthy appendix devoted to fluorescent proteins has been added that discusses the assay applications and properties of these important “reporter” molecules.

Emphasis is on discovery and R&D stages, although these assays are also important for manufacturing and QC. This report will present information on both high-content and highthroughput assays. Because it is intended to facilitate the proper choice of assay formats, this report includes many detailed comparisons between the leading possibilities for assays.

This report also emphasizes certain, of the more common, assays in biopharma. These include specific discussions of measuring compound quantity, microbial contamination, binding, enzymes, G protein-coupled receptors (GPCRs), ion channels, toxicology-pharmacology, and genetic polymorphism. It also includes emphasis on cell-based assay formats, and the most modern assay platforms, featuring miniaturization and automation.

This report presents emerging technologies for assays from the point of view of capabilities, strengths and weaknesses, and expectations for the future. The vocabulary and style of this report assume that the reader has completed at least a bachelor level education in molecular biology, biochemistry, immunology, or a similar scientific field. Experience or familiarity with assay terminology and technologies are generally not assumed, but an understanding of the business context of the assay program is important for implementing the procedures described herein.

This report does contain some technical instructions; in order to be truly useful, many tips for troubleshooting comprise specific materials and methods, however, it is not a laboratory recipe book. Detailed laboratory procedures are mostly left to development by individual labs, in response to local assay goals, policies, and requirements.

The information contained here is intended to provide knowledge of assays and assay development that normally accumulate over many years of personal experience with laboratory assay technology. This information does not supersede requirements imposed by the assay laboratory’s corporate, legal, or regulatory environment. While it cannot replace the judgment of experienced laboratory scientists and team managers, it should greatly assist such persons in their duties.

Get Full Details About This Report >>

US: 800.298.5699

Int'l: +1.240.747.3093

advertise with us

About MarketResearch.com
MarketResearch.com is an online aggregator selling over 160,000 market research reports, company profiles and country profiles from over 600 research firms. Our reports will provide you with the critical business and competitive intelligence you need for strategic planning and marketing research. Coverage includes the US, UK, Europe, Asia and global markets.

Assay Development and Technologies (2nd Edition)

Table of Contents

Abstract