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This collection seeks to elucidate the practical methods necessary for successful adjuvant development, with a particular focus on the synthesis, formulation, manufacturing, and characterization aspects involved. Beginning with an overview and a case study, the book then delves into in silico design, chemical synthesis, biosynthesis, and/or purification from natural raw materials of specific adjuvant molecules, adjuvant formulation approaches, the analytical characterization of adjuvant formulations and adjuvant-containing vaccines, as well as the biological characterization of vaccine adjuvant activity, including in vitro and in vivo approaches, to measure innate and adaptive immune responses. Written in the highly successful Methods in Molecular Biology format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Practical and authoritative, Vaccine Adjuvants: Methods and Protocols aims to facilitate vaccine adjuvant product development efforts, making them more accessible, manufacturable, and better characterized.
Sommario
Overview of Vaccine Adjuvants: Introduction, History, and Current Status.- Development of the CpG Adjuvant 1018: A Case Study.- Syntheses of Human TLR8-Specific Small-Molecule Agonists.- Semisynthesis of Analogues of the Saponin Immunoadjuvant QS-21.- QS-21 Adjuvant: Laboratory-Scale Purification Method and Formulation into Liposomes.- Purification of an Immunoadjuvant Saponin Fraction from Quillaja brasiliensis Leaves by Reverse-Phase Silica Gel Chromatography.- Biosynthetic Approaches to Squalene Production: The Case of Yeast.- In Silico Adjuvant Design and Validation.- Manufacturing Methods for Liposome Adjuvants.- Synthesis of Lymph Node-Targeting Adjuvants.- Preparing an Adjuvanted Thermoresponsive Gel Formulation for Sublingual Vaccination.- Manufacture of Oil-in-Water Emulsion Adjuvants.- Methods to Prepare Aluminum Salt-Adjuvanted Vaccines.- Production of Adjuvant-Loaded Biodegradable Particles for Use in Cancer Vaccines.- Lyophilization of Adjuvanted Vaccines: Methods for Formulation of a Thermostable Freeze Dried Product.- Stressed Stability Techniques for Adjuvant Formulations.- Particle Sizing of Nanoparticle Adjuvant Formulations by Dynamic Light Scattering (DLS) and Nanoparticle Tracking Analysis (NTA).- Quantification of Multiple Components of Complex Aluminum-Based Adjuvant Mixtures by Using Fourier Transform Infrared Spectroscopy and Partial Least Squares Modeling.- Determination of Protein Content in Alhydrogelâ-Based Vaccines by O-Phthalaldehyde Assay.- Staining and Transfer Techniques for SDS-PAGE Gels to Minimize Oil-in-Water Emulsion Adjuvant Interference.- Interactions Between Antigens and Nanoemulsion Adjuvants: Separation and Characterization Techniques.- Screening Vaccine Formulations in Fresh Human Whole Blood.- Analysis of the Innate Response to Adjuvants: Characterization of the Draining Lymph Node by Fluorescence-Activated Cell Sorting.- Assessment of Antigen-Specific Cellular Immunogenicity Using Intracellular Cytokine Staining, ELISpot, and Culture Supernatants.- Eliciting Epitope-Specific CD8 T Cell Response by Immunization with Microbial Protein Antigens Formulated with -Galactosylceramide: Theory, Practice, and Protocols.- Molecular Methods and Bioinformatic Tools for Adjuvant Characterization by High-Throughput Sequencing.
Riassunto
This collection seeks to elucidate the practical methods necessary for successful adjuvant development, with a particular focus on the synthesis, formulation, manufacturing, and characterization aspects involved. Beginning with an overview and a case study, the book then delves into
in silico
design, chemical synthesis, biosynthesis, and/or purification from natural raw materials of specific adjuvant molecules, adjuvant formulation approaches, the analytical characterization of adjuvant formulations and adjuvant-containing vaccines, as well as the biological characterization of vaccine adjuvant activity, including in vitro and in vivo approaches, to measure innate and adaptive immune responses. Written in the highly successful
Methods in Molecular Biology
format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Practical and authoritative,
Vaccine Adjuvants: Methods and Protocols
aims to facilitate vaccine adjuvant product development efforts, making them more accessible, manufacturable, and better characterized.
