Coupled Bioluminescent Assays: Methods, Evaluations, and Applications

Brings the field of coupled bioluminescence to mainstream science
This book enables readers to take full advantage of coupled bioluminescent (CB) assay techniques by demonstrating how they are used to solve real–world problems. It guides readers involved in drug discovery and biomedicine through a broad range of valuable CB techniques, including assays of kinases, phosphatases, and other enzymes; cytotoxicity measurements; and high–throughput screening of receptor modulators. In each case, CB methods are compared with competing technologies to enable informed choices. In addition, the book′s coverage of CB methods in biomonitoring; analysis of nitrate, a critical marker of explosives residue and fertilizer runoff; and nerve agent and pesticide detection supports the work of researchers involved in biodefense as well as environmental and food sciences.
Coupled Bioluminescent Assays is divided into three parts:
Background to coupled bioluminescent assays
Biomedical applications of coupled bioluminescence
Other applications of coupled bioluminescence
Highlights of coverage include:
CB cytotoxicity assays
CB assays of kinases and phosphatases
CB measurements of G protein coupled receptor activity
Assessment of nitric oxide synthase activity by CB
CB determination of bioburden and sterility
Emerging regulatory concerns
Each chapter clearly presents both underlying theory and applications. References facilitate further investigations by guiding readers to the primary literature.
CB techniques offer advantages in speed and sensitivity that fluorometric techniques generally cannot match. Bioluminescent reaction schemes are typically one to several orders of magnitude more sensitive than fluorometric assays, yet these techniques have been vastly underutilized. With this book as their guide, chemists, biochemists, and molecular biologists can take full adv antage of a broad range of available CB assays to achieve their research goals.

Spis treści:
PREFACE.
PART I BACKGROUND TO COUPLED BIOLUMINESCENT ASSAYS.
1 Introduction.
1.1 Introduction to Coupled Bioluminescent Assays.
1.2 Luminescent Technologies of the Life Sciences.
1.3 Varieties of Fluorometric Assays.
1.4 Chemiluminescence and Bioluminescence.
1.5 Common Bioluminescence Systems.
1.6 A Coupled Bioluminescent Reaction.
1.7 Summary.
2 Coupled Bioluminescent Reactions in Practice.

2.1 Principles of Coupled Bioluminescent Reactions.
2.2 Instrumentation and Equipment for Coupled Bioluminescent Assays.
2.3 Coupled Bioluminescent Assay Procedures and Precautions.
2.4 Data Handling for Coupled Bioluminescent Assays.
2.5 Comparison of Coupled Bioluminescent Assays with Other Methods.
PART II BIOMEDICAL APPLICATIONS OF COUPLED BIOLUMINESCENCE.
3 Coupled Bioluminescent Cytotoxicity Assays.
3.1 Introduction.
3.2 Membrane Integrity Assays.
3.3 A Coupled Bioluminescent Assay for Enzyme Release: The G3PDH–Release Assay.
3.4 Viability Assays.
4 The Role of Coupled Bioluminescent Assays in Kinase Screening and Study.
4.1 The Many Roles of Kinases in Biology.
4.2 Current Standard Kinase Assays.
4.3 Coupled Bioluminescent Kinase Assays.
4.4 Conclusions.
5 Coupled Bioluminescent Phosphatase Assays.
5.1 Introduction.
5.2 Phosphatases.
5.3 Contemporary Phosphatase Assay Technologies.
5.4 CB Phosphatase Assays.
5.5 Conclusions.
6 Acetylcholinesterase.
6.1 Introduction.
6.2 Established AChE Assay Methods.
6.3 Recent Developments in AChE Assay Methods.
6.4 Evaluation of Current AChE High–Throughput Screening Methods.
6.5 Coupled Bioluminescent Assays of AChE Activity.
6.6 Comparison of Coupled Bioluminescent and Other Methods of Measuring AChE Activity.
7 Measurem ent of Nitric Oxide Synthase Activity by Coupled Bioluminescence.
7.1 Introduction.
7.2 Current NOS Assays.
7.3 Conclusions.
8 The Coupled Bioluminescent Pyrophosphorolysis Assay.
8.1 Introduction.
8.2 Genetic Variation in Modern Medicine.
8.3 DNA Complementarity.
8.4 The READIT® Pyrophosphorolysis Assay.
9 Coupled Luminescent Assays of G–Protein–Coupled Receptors.

9.1 Introduction to G–Protein–Coupled Receptors.
9.2 GPCR Assay Methods.
9.3 Summary.
10 Coupled Bioluminescent Protease Assays.

10.1 Introduction.
10.2 Protease Assays.
10.3 Coupled Bioluminescent Protease Assays.
10.4 Summary.
11 Coupled Luminescent Assays Involving Aequorin.
11.1 Introduction to Aequorin.
11.2 Detection of Calcium in Practice.
12 Coupled Bioluminescent Reporter Assays.
12.1 Introduction to Reporter Assays.
12.2 Luciferases as Reporters of Promoter Activities.
12.3 Aequorin as a Reporter Enzyme.
12.4 Vectors for Use in Reporter Assays.
12.5 Summary.
13 Coupled Bioluminescent Assays: Regulatory Concerns.

13.1 Introduction.
13.2 Regulatory Aspects of Assay Development.
13.3 Summary.
PART III OTHER APPLICATIONS OF COUPLED BIOLUMINESCENCE.
14 Coupled Bioluminescent Determination of Bioburden and Sterility.
14.1 Introduction.
14.2 Rapid Methods of Bioburden and Sterility Assessment.
15 Environmental Applications of Coupled Bioluminescent Assays.
15.1 Introduction.
15.2 Current Methods for Environmental Monitoring of Water Quality.
15.3 Methods of Monitoring Stream Water and Lake Water.
15.4 Methods of Monitoring Drinking Water.
Appendix A: One–Letter Amino Acid Abbreviations.
Glossary.
Bibliography.
Index.

Nota biograficzna:
Michael J. Corey, PHD, is a consulting biochemist and a leading authority on coupled bioluminescent assays. A former head of assay development in biotechnology, he is a founding member of the Scientific Advisory Board of Cell Technology, Inc. Dr. Corey holds key patents in the field of coupled bioluminescence.

Okładka tylna:
Brings the field of coupled bioluminescence to mainstream science
This book enables readers to take full advantage of coupled bioluminescent (CB) assay techniques by demonstrating how they are used to solve real–world problems. It guides readers involved in drug discovery and biomedicine through a broad range of valuable CB techniques, including assays of kinases, phosphatases, and other enzymes; cytotoxicity measurements; and high–throughput screening of receptor modulators. In each case, CB methods are compared with competing technologies to enable informed choices. In addition, the book′s coverage of CB methods in biomonitoring; analysis of nitrate, a critical marker of explosives residue and fertilizer runoff; and nerve agent and pesticide detection supports the work of researchers involved in biodefense as well as environmental and food sciences.
Coupled Bioluminescent Assays is divided into three parts:
Background to coupled bioluminescent assays
Biomedical applications of coupled bioluminescence
Other applications of coupled bioluminescence
Highlights of coverage include:
CB cytotoxicity assays
CB assays of kinases and phosphatases
CB measurements of G protein coupled receptor activity
Assessment of nitric oxide synthase activity by CB
CB determination of bioburden and sterility
Emerging regulatory concerns
Each chapter clearly presents both underlying theory and applications. References facilitate further investigations by guiding readers to the primary literature.
CB techniques offer advantages in speed and sensitivity that fluorometric techniques generally cannot match. Bioluminescent reaction schemes are typically one to several orders of magnitude more sensitive than fluorometric assays, yet these techniques have been vastly underutilized. With this book as their guide, chemists, biochemists, and molecular biologists can take full advantage of a broad range of available CB assays to achieve their research goals.