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.