Molecular Diagnosis of Infectious Diseases

Front Cover
Jochen Decker, Udo Reischl
Springer Science & Business Media, 2004 - Medical - 464 pages
In this second edition of a much-acclaimed classic laboratory manual, Molecular Diagnosis of Infectious Diseases, leading experts from academia and industry describe cutting-edge methods for the protein-based diagnosis of infectious diseases. Explaining the latest developments in genomics, proteomics, bioinformatics, biosensors, high-throughput devices, and recombinant technology, the authors demonstrate the power of these new methodologies in both established and novel procedures by applying them successfully to the identification and characterization of valuable diagnostic markers, immunomodulatory components, epitope mapping, the production and purification of recombinant antigens, and their use in diagnostic reagents in immunological assays. Basic problems of serological laboratory diagnostics and the use of immunoproteomics for discovering target proteins are also addressed. Each readily reproducible protocol is described in step-by-step detail and includes a background introduction, equipment and reagent lists, tips on troubleshooting and avoiding known pitfalls, and extensive references. Comprehensive and up-to-date, Molecular Diagnosis of Infectious Diseases, Second Edition offers novice investigators an easy entry into the field and experienced researchers a broad array of time saving tools that will not only elucidate new principles, but also greatly facilitate the development and successful application of rapid diagnostic methods.
 

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Contents

Foreword
v
Preface
vii
Contributors
xii
PROTEINBASED IDENTIFICATION OF ANTIGENS
2
Proteomic Approaches to Antigen Discovery
3
Immunoproteomics
19
Immunoprecipitation and Blotting The Visualization of Small Amounts of Antigens Using Antibodies and Lectins
33
DNABASED IDENTIFICATION OF ANTIGENS
47
CodonOptimized Genes that Enable Increased Heterologous Expression in Mammalian Cells and Elicit Efficient Immune Responses in Mice after Vac...
197
PURIFICATION MODIFICATION AND RENATURATION OF RECOMBINANT PROTEINS
211
Purification and Immunological Characterization of Recombinant Antigens Expressed in the Form of Insoluble Aggregates Inclusion Bodies
213
Purification of Recombinant Proteins with High Isoelectric Points
225
Refolding of Inclusion Body Proteins
239
SmallMoleculeProtein Conjugation Procedures
255
CHARACTERIZATION OF RECOMBINANT PROTEINS
267
Structural Characterization of Proteins and Peptides
269

Representational Difference Analysis of cDNA
49
Microarray Data Analysis and Mining
67
Expression Cloning
91
IDENTIFICATION OF EPITOPES AND IMMUNOMODULATORY COMPONENTS
107
Determination of Epitopes by Mass Spectrometry
109
Identification of TCell Epitopes Using ELISpot and Peptide Pool Arrays
121
Viruslike Particles A Novel Tool for the Induction and Monitoring of Both THelper and Cytotoxic TLymphocyte Activity
133
Application of SingleCell Cultures of Mouse Splenocytes as an Assay System to Analyze the Immunomodulatory Properties of Bacterial Components
159
EXPRESSION OF RECOMBINANT PROTEINS
177
HighThroughput Expression and Purification of 6xHisTagged Proteins in a 96Well Format
179
Production of Antigens in Chlamydomonas reinhardtii Green Microalgae as a Novel Source of Recombinant Proteins
191
Determination of Kinetic Data Using Surface Plasmon Resonance Biosensors
299
Affinity Measurements of Biological Molecules by a Quartz Crystal Microbalance QCM Biosensor
321
EVALUATION OF RECOMBINANT PROTEINS IN IMMUNOLOGICAL TEST SYSTEMS
331
Solid Supports in EnzymeLinked Immunosorbent Assay and Other SolidPhase Immunoassays
333
Design and Preparation of Recombinant Antigens as Diagnostic Reagents in SolidPhase Immunosorbent Assays
373
Basic Problems of Serological Laboratory Diagnosis
393
Molecular Diagnostics Resources on the Internet
429
RECOMBINANT RECEPTOR MOLECULES
445
Cloning SingleChain Antibody Fragments scFv from Hybridoma Cells
447
Index
459
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