A comprehensive and up-to-date set of basic and advanced methods that illuminates the principles of molecular diagnostics and allows researchers to establish assay systems tailored to their specific needs. This unique collection has been thoroughly updated and expanded and covers the latest strategies for identifying novel diagnostic markers, epitope mapping, and the production and purification of recombinant antigens and their use as diagnostic reagents in immunological assays. This new edition places significant emphasis on bioinformatics and proteomics and the characterization and modification of proteins. It also includes promising techniques based on recent developments in antibody engineering and on the use of biosensors. Rather than focusing on particular infectious agents, this book focuses on the techniques that can be used and tailored to accomplish any kind of molecular diagnostic application in the clinical laboratory, thus making it an essential book for all laboratory disagnosticians.
Contents:
1. Identification of Novel Diagnostic Markers by Differential Display
Peng Liang; Fellan Wang; Welmin Zhu; Robert O'Connell; Lidia Averboukh
pp. 3-13
2. lmmunoprecipitation
Kari Johansen; Lennart Svensson
pp. 15-28
3. RTPCR Methods and Applications
Paul Siebert
pp. 29-54
4. Quantitative RT-PCR
Paul Siebert
pp. 55-80
5. Cloning cDNA Ends Using RACE
Yue Zhang; Michael Frohman
pp. 81-106
6. A Method for the Rapid Identification of Epitopes and Other Functional Peptide Domains
Robert Mierendorf; Beth Hammer; Robert Novy
pp. 107-124
7. Mapping of Linear B-Cell Epitopes on Viral Polypeptides by Multiple Peptide Synthesis and Fine Tuning Sensitivity and Specificity of the Identified Peptide Antigens for Application in Virus Diagnosis
Ulla Wienhues; Wolfgang Kraas; Christoph Seidel; Hans Ihlenfeldt; Günther Jung
pp. 125-142
8. Amplification of Genomic DNA by PCR
Bruno Frey
pp. 143-156
9. PCR-Based Cloning and Subsequent Expression of Antigenic Proteins in Escherichia coli
Udo Reischl
pp. 157-168
10. Application of TEV Protease in Protein Production
Deborah Polayes; T. Parks; Stephen Johnston; William Dougherty
pp. 169-184
Table of Contents
I. Recombinant Proteins and Peptides.
Identification of Novel Diagnostic Markers by Differential Display
Peng Liang, Feilan Wang, Weimin Zhu, Robert P. O'Connell, and Lidia Averboukh
Immunoprecipitation
Kari Johansen and Lennart Svensson
RT-PCR Methods and Applications
Paul D. Siebert
Quantitative RT-PCR
Paul D. Siebert
Cloning cDNA Ends Using RACE
Yue Zhang and Michael A. Frohman
A Method for the Rapid Identification of Epitopes and Other Functional Peptide Domains
Robert C. Mierendorf, Beth Hammer, and Robert E. Novy
Mapping of Linear B-Cell Epitopes on Viral Polypeptides by Multiple Peptide Synthesis and Finetuning Sensitivity and Specificity of the Identified Peptide Antigens for Application in Virus Diagnosis
Ulla Wienhues, Wolfgang Kraas, Christoph Seidel, Hans Georg Ihlenfeldt, and Günther Jung
Amplification of Genomic DNA by PCR
Bruno Frey
PCR-Based Cloning and Subsequent Expression of Antigenic Proteins in Escherichia coli
Udo Reischl
Application of TEV Protease in Protein Production
Deborah A. Polayes, T. Dawn Parks, Stephen A. Johnston, and William G. Dougherty
Stable Transfected Drosophila Schneider-2 Cells as a Novel Tool to Produce Recombinant Antigens for Diagnostic, Therapeutic, and Preventive Purposes
Ludwig Deml and Ralf Wagner
Use of Recombinant Antigens as Diagnostic Reagents in Enzyme-Linked Immunosorbent Assays
Alan Warnes, Anthony R. Fooks, and John R. Stephenson
Generation of Recombinant Baculovirus DNA in E. coli Using a Baculovirus Shuttle Vector
Valentina C. Ciccarone, Deborah A. Polayes, and Verne A. Luckow
The SFV Gene Expression System
Valentina C. Ciccarone, Joel A. Jessee, and Peter Liljeström
Expression and Purification of Recombinant Proteins Using the pET System
Robert C. Mierendorf, Barbara B. Morris, Beth Hammer, and Robert E. Novy
Procedures for the Analysis and Purification of His-Tagged Proteins
Richard E. Kneusel, Joanne Crowe, Melanie Wulbeck, and Joachim Ribbe
Detection and Immobilization of Proteins Containing the 6xHis Tag
Richard E. Kneusel, Melanie Wulbeck, and Joachim Ribbe
The Use of Recombinant Fusion Proteases in the Affinity Purification of Recombinant Proteins
Louis E. C. Leong
Purification and Immunological Characterization of Recombinant Antigens Expressed in the Form of Insoluble Aggregates (Inclusion Bodies)
Udo Reischl
Purification of Recombinant Proteins Having High Isoelectric Points
Raffaele A. Calogero and Anna Aulicino
Immunoblotting
Michael Kirschfink and Peter Terness
Diagnostic Application of a Multiple-Channel Immunoblot Apparatus
Shaun D. Black
Immobilization of Immunoreactants on Solid Phases for ELISAs
Richard Decker and David Leahy
Nonradioactive Labeling and Immunodetection of Recombinant Proteins
Robert Oberfelder
Flow Cytometry
Volker Daniel
Basic Problems of Serological Laboratory Diagnosis
Walter Fierz
Molecular Diagnostics Resources on the Internet
Kevin J. O'Donnell
II. Antibodies.
Monoclonal Antibody-Based Immunoassays
Peter Appleby and Udo Reischl
Biosensor-Based Methods in Clinical Diagnosis
Franz Aberl and Conrad Kößlinger
The Quartz Crystal Microbalance (QCM) as an Immunosensor
Franz Aberl, Conrad Kößlinger, and Hans Wolf
Biomolecular Interaction Analysis Strategies for Antibody and Antigen Characterization
Uwe W. Roder and Francis Markey
Recent Developments in Antibody Engineering
Melvyn Little, Stefan Dübel, Sergey Kipriyanov, and Frank Breitling
Cloning and Expression of Single Chain Fragments (scFv) from Mouse and Rat Hybridomas
Frank Breitling and Stefan Dübel
Production of a Human Antibody Library in the Phage-Display Vector pSEX81
Martin Welschof, Melvyn Little, and Heinz Dörsam
Screening of Phage-Displayed Antibody Libraries
Heinz Dörsam, Michael Braunagel, Christian Kleist, Daniel Moynet, and Martin Welschof
Purification, Characterization, and Biotinylation of Single-Chain Antibodies
Sergey M. Kipriyanov
Index.
Un jeu complet(compréhensif) et à jour des méthodes de **** et avancées qui illuminent les principes de diagnostic moléculaire et permet aux chercheurs d'établir des systèmes d'essai façonnés à leurs besoins spécifiques. Cette collection(ramassage) unique a été à fond mise à jour et étendue et couvre les dernières stratégies pour l'identification de nouveaux marqueurs diagnostiques, epitope la configuration et la production et la purification de recombinant antigens et leur utilisation comme des réactifs diagnostiques dans des essais immunologiques. Cette nouvelle édition place l'accent significatif sur bioinformatics et proteomics et la caractérisation et la modification de protéines. Il inclut aussi des techniques prometteuses basées sur des événements récents dans l'ingénierie d'anticorps et sur l'utilisation de biosensors. Plutôt que des agents se concentrant particuliers infectieux, ces centres de livre sur les techniques qui peuvent être employées et façonnés pour accomplir n'importe quelle sorte de demande(application) moléculaire diagnostique dans le laboratoire d'analyses médicales, le faisant ainsi un livre essentiel pour tout le laboratoire disagnosticians.
Contenu :
1. Identification de Nouveaux Marqueurs Diagnostiques par Exposition Différentielle
Peng Liang; Fellan Wang; Welmin Zhu; Robert O'Connell; Lidia Averboukh
Pp. 3-13
2. Lmmunoprecipitation
Kari Johansen; Lennart Svensson
Pp. 15-28
3. RTPCR Méthodes et Demandes(applications)
Paul Siebert
Pp. 29-54
4. RT-PCR quantitatif
Paul Siebert
Pp. 55-80
5. Clonage(multiplication) cDNA Fins Employant COURSE(RACE)
Yue Zhang; Michel Frohman
Pp. 81-106
6. Une Méthode pour l'Identification Rapide d'Epitopes et d'Autres Domaines de Peptide Fonctionnels
Robert Mierendorf; Beth Hammer; Robert Novy
Pp. 107-124
7. Configuration de B-cellule Linéaire Epitopes sur Polypeptides Viraux par Synthèse de Peptide Multiple et Sensibilité de Réglage d'accord Excellente et Spécificité du Peptide Identifié Antigens pour Demande(application) dans Diagnostic Viral
Ulla Wienhues; Wolfgang Kraas; Christoph Seidel; Hans Ihlenfeldt; Günther Jung
Pp. 125-142
8. Amplification d'ADN Génomique par PCR
Bruno Frey
Pp. 143-156
9. Clonage(multiplication) PCR-BASÉ et Expression Suivante de Protéines Antigenic dans Escherichia coli
Udo Reischl
Pp. 157-168
10. Demande(application) de TEV Protease dans Production de Protéine
Deborah Polayes; T. Parcs; Stéphane Johnston; William Dougherty
Pp. 169-184
المواضيع المتشابهه
Molecular Diagnosis of Infectious Diseases
العرض العادي
