Vaccine research and development is advancing at an unprecedented pace, with an increasing emphasis on rational design based upon a fundamental und- standing of the underlying molecular mechanisms. The aim of this volume is to provide a selection of contemporary protocols that will be useful to both novice and advanced practitioner alike. The variety of procedures required to design, develop, produce, and assess a vaccine is immense and covers aspects of ch- istry, biochemistry, molecular biology, cell biology, and immunology. No single volume can hope to cover these topics exclusively. Rather, here we attempt to provide a methods sourcebook focusing on hands-on practical advice. Comp- mentary and background information may be found in other volumes in the Me- ods in Molecular Medicine series. Of particular interest are volumes on Dendritic Cell Protocols, Interleukin Protocols, Vaccine Adjuvants, and DNA Vaccines. Since the publication of the first edition of Vaccine Protocols there have been major advances, particularly in the areas of bacterial genomics, antig- specific T-cell quantification, genetic manipulation of vaccine vectors, the h- nessing of natural molecules concerned with the regulation of immune responses, and the burgeoning field of DNA vaccinology. Hence, the extensive revision of this edition with new chapters on live viral vaccine vectors, atte- ated bacterial vectors, immunomodulators, MHC-peptide tetrameric complexes, and the identification of vaccine candidates by genomic analysis. Additionally, chapters from the first edition have been updated to accommodate state-of-t- art methods in vaccinology.

Includes supplementary material: sn.pub/extras

Klappentext
This extensively revised, updated, and expanded second edition of a classic laboratory manual applies new understanding of molecular mechanisms to the rational design and development of vaccines. Here the working researcher will find readily reproducible techniques for the development and production of live attenuated vaccines through the genetic manipulation of viruses and bacteria, vaccine vectors or inactivated toxins, and the production of synthetic peptides and conjugate vaccines. In addition, there are chapters describing more general techniques for vaccine formulation, delivery, and assessment of immune responses, and review discussions of scale-up to manufacture, vaccine quality assurance, and clinical trials. New to this second edition are cutting-edge chapters on two different viral vectors (Vaccinia and Semliki Forest), lactic acid bacteria for delivery of foreign antigens, incorporation of immunomodulators into plasmid DNA vaccines, construction of tetramer complexes to detect CMI responses, assessment of functional antibody responses, and the use of complete genome sequences for vaccine design. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes a background introduction outlining the principle behind the technique, equipment and reagent lists, tips on trouble shooting and avoiding known pitfalls, and, where needed, a discussion of the interpretation and use of results.
State-of-the-art and highly practical, Vaccine Protocols, Second Edition, provides essential information for those seeking to produce improved or novel vaccines in the rapidly evolving field of vaccine research and development.

Inhalt
Overview of Vaccines.- Temperature-Sensitive Mutant Vaccines.- Live Viral Vectors.- Live Viral Vectors.- Live Viral Vectors.- Development of Attenuated Salmonella Strains That Express Heterologous Antigens.- Expression and Delivery of Heterologous Antigens Using Lactic Acid Bacteria.- Synthetic Peptides.- Genetic Detoxification of Bacterial Toxins.- Preparation of Polysaccharide-Conjugate Vaccines.- Adjuvant Formulations for Experimental Vaccines.- Incorporation of Immunomodulators into Plasmid DNA Vaccines.- Microencapsulation of Vaccine Antigens.- Lyophilization of Vaccines.- Stimulation of Mucosal Immunity.- Induction and Detection of T-Cell Responses.- Construction of MHC Class I-Peptide Tetrameric Complexes for Analysis of T-Cell-Mediated Immune Responses.- Assessment of Functional Antibody Responses.- The Use of Complete Genome Sequences in Vaccine Design.- Severe Combined Immunodeficient (SCID) Mice in Vaccine Assessment.- Clinical Trials.- Assuring the Quality and Safety of Vaccines.- DNA Vaccination.- From Vaccine Research to Manufacture.