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DNA Vaccines Methods and Protocols 1st Edition by Douglas Lowrie, Robert Whalen ISBN 9781592596881 159259688X

Name: DNA Vaccines Methods and Protocols 1st Edition by Douglas Lowrie, Robert Whalen ISBN 9781592596881 159259688X
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Authors:Douglas B. Lowrie , Series:Forensic [1] , Tags:Forensic Medicine and Toxicology , Author sort:Lowrie, Douglas B. , Languages:Languages:eng , Publisher:Humana Press , Comments:Comments:Current technologies for the purification of supercoiled plasmids are limited.The use of cesium chloride gradients in the presence of ethidium bromideis time consuming, labor intensive, requires the use of known mutagens and isnot conducive to large scale. As a result, first-generation high-performanceliquid chromatography (HPLC) methods based on anion-exchange and sizeexclusion have been developed but are difficult to accommodate production atlarge scale and still result in compromised purity (1,2). The success of DNAvaccines in animal models and the initiation of human trials (3,4) has led to aneed to increase the level of supercoiled plasmid purity as well as the methodologyutilized to produce these plasmids at large scale. Several parameters ofthe purification process need to be addressed:

SKU: EB-10990 Category: eBook PDF Tags: DNA Vaccines Methods, Douglas Lowrie, Protocols, Robert Whalen

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ISBN 10: 159259688X
ISBN 13: 9781592596881
Author: Douglas Lowrie, Robert Whalen

The field of DNA vaccines has undergone explosive growth in the last few years. As usual, some historical precursors of this approach can be d- cerned in the scientific literature of the last decades. However, the present state of affairs appears to date from observations made discreetly in 1988 by Wolff, Malone, Felgner, and colleagues, which were described in a 1989 patent and published in 1990. Quite surprisingly, they showed that genes carried by pure plasmid DNA and injected in a saline solution, hence the epithet “naked DNA,” could be taken up and expressed by skeletal muscle cells with a low but reproducible frequency. Such a simple methodology was sure to spawn many applications. In a separate and important line of experimentation, Tang, De Vit, and Johnston announced in 1992 that it was indeed possible to obtain humoral immune responses against proteins encoded by DNA delivered to the skin by a biolistic device, which has colloquially become known as the “gene gun. ” The year 1993 saw the publication of further improvements in the me- ods of naked DNA delivery and, above all, the first demonstrations by several groups of the induction of humoral and cytotoxic immune responses to viral antigens expressed from injected plasmid DNA. In some cases, protection against challenge with the pathogen was obtained. The latter result was – questionably the touchstone of a method of vaccination worthy of the name.

DNA Vaccines Methods and Protocols 1st Edition Table of contents:

Introduction to DNA Vaccines
- Overview of DNA Vaccine Technology
- History and Development of DNA Vaccines
- Applications of DNA Vaccines in Veterinary and Human Medicine
Basic Concepts in DNA Vaccination
- Mechanism of Action of DNA Vaccines
- Immune Responses Induced by DNA Vaccines
- Advantages and Challenges of DNA Vaccines
Designing DNA Vaccines
- Choice of Antigen and Plasmid Vector
- Codon Optimization and Adjuvant Incorporation
- Strategies for Enhancing Immune Responses
Preparation of DNA Vaccine Plasmids
- Cloning of Antigen Genes into Vaccine Vectors
- Plasmid Amplification and Purification
- Quality Control and Validation of Plasmids
Gene Delivery Systems
- Electroporation Techniques
- Particle Bombardment (Gene Gun)
- Liposome and Lipid-Mediated Delivery
- Viral Vectors for DNA Vaccine Delivery
Immunization Protocols
- DNA Vaccine Administration Methods (Intramuscular, Subcutaneous, Intradermal)
- Dosing Regimens and Frequency of Immunization
- Use of Adjuvants to Enhance Immune Responses
Assessing Immune Responses to DNA Vaccines
- ELISA for Antibody Detection
- Cytokine Assays and Flow Cytometry for T-cell Responses
- In vivo Models for Testing Immune Responses
- Efficacy of DNA Vaccines in Animal Models
Preclinical and Clinical Testing of DNA Vaccines
- Design of Animal Studies for DNA Vaccine Testing
- Safety and Toxicity Testing
- Transitioning from Preclinical to Clinical Trials
- Regulatory Considerations for DNA Vaccine Approval
Applications of DNA Vaccines
- DNA Vaccines for Infectious Diseases
- Cancer Immunotherapy with DNA Vaccines
- DNA Vaccines for Allergies and Autoimmune Diseases
Troubleshooting DNA Vaccine Experiments
- Common Problems in DNA Vaccine Preparation and Delivery
- Optimizing Transfection Efficiency
- Interpretation of Immunological Assays
Emerging Trends and Future Directions
- Advancements in DNA Vaccine Technology
- DNA Vaccines and Next-Generation Sequencing
- Personalized DNA Vaccines and Precision Medicine
- Nanoparticle Delivery Systems for DNA Vaccines