Molecular Diagnosis of Infectious Diseases

This second edition of a classic laboratory manual describes 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 apply these new methodologies successfully to the identification and characterization of valuable diagnostic markers, immunomodulatory components, epitope mapping, the production and purification of recombinant antigens, as well as to diagnostic reagents in immunological assays.

Foreword

v

Preface

vii

Contributors

xiii

Part I. Protein-Based Identification of Antigens

Proteomic Approaches to Antigen Discovery

3

(16)

Karen M. Dobos

John S. Spencer

Ian M. Orme

John T. Belisle

Immunoproteomics

19

(14)

Alexander Krah

Peter R. Jungblut

Immunoprecipitation and Blotting: The Visualization of Small Amounts of Antigens Using Antibodies and Lectins

33

(16)

Stephen Thompson

Part II. Dna-Based Identification of Antigens

Representational Difference Analysis of cDNA

49

(18)

Lucas D. Bowler

Microarray Data Analysis and Mining

67

(24)

Silvia Saviozzi

Giovanni Iazzetti

Enrico Caserta

Alessandro Guffanti

Raffaele A. Calogero

Expression Cloning

91

(18)

Michael J. Lodes

Davin C. Dillon

Raymond L. Houghton

Yasir A. W. Skeiky

Part III. Identification of Epitopes and Immunomodulatory Components

Determination of Epitopes by Mass Spectrometry

109

(12)

Christine Hager-Braun

Kenneth B. Tomer

Identification of T Cell Epitopes Using ELISpot and Peptide Pool Arrays

121

(12)

Timothy W. Tobery

Michael J. Caulfield

Virus-like Particles: A Novel Tool for the Induction and Monitoring of Both T-Helper and Cytotoxic T-Lymphocyte Activity

133

(26)

Ludwig Deml

Jens Wild

Ralf Wagner

Application of Single-Cell Cultures of Mouse Splenocytes as an Assay System to Analyze the Immunomodulatory Properties of Bacterial Components

159

(20)

Ludwig Deml

Michael Aigner

Alexander Eckhardt

Jochen Decker

Norbert Lehn

Wulf Schneider-Brachert

Part IV. Expression of Recombinant Proteins

High-Throughput Expression and Purification of 6xHis- Tagged Proteins in a 96-Well Format

179

(12)

Jutta Dress

Jason Smith

Frank Schafer

Kerstin Steinert

Production of Antigens in Chlamydomonas reinhardtii: Green Microalgae as a Novel Source of Recombinant Proteins

191

(6)

Markus Fuhrmann

Codon-Optimized Genes that Enable Increased Heterologous Expression in Mammalian Cells and Elicit Efficient Immune Responses in Mice after Vaccination of Naked DNA

197

(16)

Marcus Graf

Ludwig Deml

Ralf Wagner

Part V. Purification, Modification, and Renaturation of Recombinant Proteins

Purification and immunological Characterization of Recombinant Antigens Expressed in the Form of Insoluable Aggregates (Inclusion Bodies)

213

(12)

Udo Reischl

Purification of Recombinant Protins with High Isoelectric Points

225

(14)

Raffaele A. Calogero

Anna Aulicino

Refolding of Inclusion Body Proteins

239

(16)

Marcus Mayer

Johannes Buchner

Small-Molecule-Protein Conjugation Procedures

255

(14)

Stephen Thompson

Part VI. Characterization of Recombinant Proteins

Structural Characterization of Proteins and Peptides

269

(30)

Rainer Deutzmann

Determination of Kinetic Data Using Surface Plasmon Resonance Biosensors

299

(22)

Claudia Hahnefeld

Stephan Drewianka

Friedrich W. Herberg

Affinity Measurements of Biological Molecules by a Quartz Crystal Microbalance (QCM) Biosensor

321

(12)

Uwe Schaible

Michael Liss

Elke Prohaska

Jochen Decker

Karin Stadtherr

Hans Wolf

Part VII. Evaluation of Recombinant Proteins in Immunological Test Systems

Solid Supports in Enzyme-Linked Immunosorbent Assay and Other Solid-Phase Immunoassays

333

(40)

John E. Butler

Design and Preparation of Recombinant Antigens as Diagnostic Reagents in Solid-Phase Immunosorbent Assays

373

(20)

Alan Warnes

Anthony R. Fooks

John R. Stephenson

Basic Problems of Serological Laboratory Diagnosis

393

(36)

Walter Fierz

Molecular Diagnostics Resources on the Internet

429

(18)

Larry Winger

Part VIII. Recombinant Receptor Molecules

Cloning Single-Chain Antibody Fragments (scFv) from Hybridoma Cells

447

(12)

Lars Toleikis

Olaf Broaders

Stefan Dubel

Index

459

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Molecular Diagnosis of Infectious Diseases

Summary

Table of Contents

Supplemental Materials

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Molecular Diagnosis of Infectious Diseases

Summary

Table of Contents

Supplemental Materials

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