Assay Development: Fundamentals and Practices

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John Wiley & Sons, Jun 25, 2010 - Science - 450 pages
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Essential principles and practice of assay development

The first comprehensive, integrated treatment of the subject, Assay Development: Fundamentals and Practices covers the essentials and techniques involved in carrying out an assay project in either a biotechnology/drug discovery setting or a platform setting.

Rather than attempting comprehensive coverage of all assay development technologies, the book introduces the most widely used assay development technologies and illustrates the art of assay development through a few commonly encountered biological targets in assay development (e.g., proteases, kinases, ion channels, and G protein-coupled receptors). Just enough biological background for these biological targets is provided so that the reader can follow the logics of assay development. Chapters discuss:

  • The basics of assay development, including foundational concepts and applications

  • Commonly used instrumental methods for both biochemical assays and cell-based assays

  • Assay strategies for protein binding and enzymatic activity

  • Cell-based assays

  • High-throughput screening

An in-depth study of the now popular Caliper's off-chip kinase assay provides an instructive, real-world example of the assay development process.

 

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Contents

1 INTRODUCTION TO ASSAY DEVELOPMENT
1
2 MEASUREMENT AND INSTRUMENTATION
27
3 FUNDAMENTAL PRINCIPLES OF ASSAY DEVELOPMENT WITH ISOLATED PROTEINS
69
4 SEPARATIONBASED TECHNIQUES IN BIOASSAYS
105
5 GENERAL PROTEIN BINDING ASSAY FORMATS
125
6 FUNCTIONAL ASSAYS WITH ISOLATED PROTEASES
155
7 FUNCTIONAL ASSAYS FOR PROTEIN KINASES
181
8 FUNDAMENTAL PRINCIPLES OF CELLBASED ASSAYS
213
9 FUNCTIONAL ION CHANNEL ASSAYS
239
10 ASSAYS WITH GPCRs
265
11 ASSAYS BASED ON INTEGRATED CELL SYSTEM PROPERTIES
289
12 HIGHCONTENT CELLBASED ASSAY WITH OPTICAL IMAGING TECHNIQUES
307
13 HIGHTHROUGHPUT SCREENING
321
14 CASE STUDY DEVELOPMENT OF A MICROFLUIDICBASED KINASE ASSAY PLATFORM
371
INDEX
423
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About the author (2010)

Ge Wu, PhD, is Director of Assay Development at Iris International. He obtained his doctorate in biochemistry from UCLA, performed postdoctoral work at Harvard Medical School, and was a member of the faculty at Cornell University Medical College. Prior to joining Iris International, he worked at Merck & Co., Caliper Technologies, Amphora Discovery, and FivePrime Therapeutics. He is the founding Director of Assay Development and Screening at FivePrime Therapeutics. Dr. Wu is a leading figure in assay development and screening in biopharmaceutical discovery using primary cell assay and the author of an acclaimed review in this subject.

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